Compounds having β2 adrenergic receptor agonist and muscarinic receptor antagonist activity

ABSTRACT

This invention provides compounds of formula I: 
     
       
         
         
             
             
         
       
         
         
           
             wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8a , R 8b , W, a and b are as defined in the specification, or a pharmaceutically acceptable salt or solvate or stereoisomer thereof. The compounds of this invention possess both β 2  adrenergic receptor antagonist and muscarinic receptor antagonist activity. Accordingly, such compounds are expected to be useful as therapeutic agents for treating pulmonary disorders, such as chronic obstructive pulmonary disease and asthma.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.60/601,781, filed on Aug. 16, 2004; the entire disclosure of which isincorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to novel compounds having β₂ adrenergicreceptor agonist and a muscarinic receptor antagonist activity. Thisinvention also relates to pharmaceutical compositions comprising suchcompounds, processes and intermediates for preparing such compounds andmethods of using such compounds to treat pulmonary disorders.

2. State of the Art

Pulmonary disorders, such as asthma and chronic obstructive pulmonarydisease (COPD), are commonly treated with bronchodilators. One class ofbronchodilator in widespread use consists of β₂ adrenergic receptor(adrenoceptor) agonists, such as albuterol, formoterol and salmeterol.These compounds are generally administered by inhalation. Another classof bronchodilator consists of muscarinic receptor antagonists(anticholinergic compounds), such as ipratropium and tiotropium. Thesecompounds are also typically administered by inhalation.

Pharmaceutical compositions containing both a β₂ adrenergic receptoragonist and a muscarinic receptor antagonist are also known in the artfor use in treating pulmonary disorders. For example, U.S. Pat. No.6,433,027 discloses medicament compositions containing a muscarinicreceptor antagonist, such as tiotropium bromide, and a β₂ adrenergicreceptor agonist, such as formoterol fumarate.

Although compounds having either β₂ adrenergic receptor agonist ormuscarinic receptor antagonist activity are known, no compound havingboth β₂ adrenergic receptor agonist and muscarinic receptor antagonistactivity has been previously disclosed. Compounds possessing both β₂adrenergic receptor agonist and muscarinic receptor antagonist activityare highly desirable since such bifunctional compounds would providebronchodilation through two independent modes of action while havingsingle molecule pharmacokinetics. Moreover, a single bifunctionalcompound would be simpler to formulate for therapeutic uses compared totwo separate compounds and would be easier to co-formulate with othertherapeutic agents to create triple therapy combinations.

SUMMARY OF THE INVENTION

The present invention provides novel compounds that have been found topossess both β₂ adrenergic receptor agonist and muscarinic receptorantagonist activity. Such compounds are expected to be useful astherapeutic agents for treating pulmonary disorders.

Accordingly, in one of its composition aspects, the present inventionprovides a compound of formula I:

wherein

W represents O or NW^(a); where W^(a) is hydrogen or (1-4C)alkyl;

R¹ is (6-10C)aryl, (2-9C)heteroaryl containing from 1 to 4 heteroatomsindependently selected from oxygen, nitrogen and sulfur or(3-7C)cycloalkyl; wherein the aryl, heteroaryl or cycloalkyl group isunsubstituted or substituted with from 1 to 3 substituents independentlyselected from (1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl,(3-6C)cycloalkyl, (6-10C)aryl, (2-9C)heteroaryl, (3-5C)heterocyclic,cyano, halo, —OR^(1a), —C(O)OR^(1b), —SR^(1c), —S(O)R^(1d),—S(O)₂R^(1e), —NR^(1f)R^(1g), —C(O)R^(1h), —NR^(1i)C(O)OR^(1j),—OC(O)NR^(1k)R^(1l), —NR^(1m)C(O)R^(1n), —C(O)NR^(1o)R^(1p) and—NHS(O)₂R^(1q); where each of R^(1a), R^(1b), R^(1c), R^(1d), R^(1e),R^(1f), R^(1g), R^(1h), R^(1i), R^(1j), R^(1k), R^(1l), R^(1m), R^(1n),R^(1o), R^(1p) and R^(1q) is independently selected from hydrogen,(1-4C)alkyl or phenyl-(1-4C)alkyl; and where each (3-6C)cycloalkyl,(6-10C)aryl, (2-9C)heteroaryl and (3-5C)heterocyclic group isunsubstituted or substituted with from 1 to 3 substituents selected from(1-4C)alkyl, halo and —OR^(1a); and where each alkyl group present in R¹is unsubstituted or substituted with from 1 to 3 fluoro substituents;

each R² is independently selected from (1-4C)alkyl, (2-4C)alkenyl,(2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(2a), —C(O)OR^(2b),—SR^(2c), —S(O)R^(2d), —S(O)₂R^(2e) and —NR^(2f)R^(2g); where each ofR^(2a), R^(2b), R^(2c), R^(2d), R^(2e), R^(2f) and R^(2g) isindependently selected from hydrogen, (1-4C)alkyl or phenyl-(1-4C)alkyl;and where each alkyl group present in R² is unsubstituted or substitutedwith from 1 to 3 fluoro substituents;

R³ is selected from hydrogen, (1-6C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl,or (3-6C)cycloalkyl, where the alkyl group is unsubstituted orsubstituted with from 1 to 3 fluoro substituents;

R⁴ represents a divalent hydrocarbon group containing from 4 to 28carbon atoms and optionally containing from 1 to 10 heteroatoms selectedindependently from halo, oxygen, nitrogen and sulfur, provided that thenumber of contiguous atoms in the shortest chain between the twonitrogen atoms to which R⁴ is attached is in the range of from 4 to 16;

R⁵ represents hydrogen or (1-4C)alkyl;

R⁶ is —N(R^(6a))C(O)R^(6b) or —CR^(6c)R^(6d)OR^(6e) and R⁷ is hydrogen;or R⁶ and R⁷ together form —N(R^(7a))C(O)C(R^(7b))═C(R^(7c))—,—C(R^(7d))═C(R^(7e))C(O)N(R^(7e))C(O)N(R^(7f))—,—N(R^(7g))C(O)CR^(7h)R^(7i)—CR^(7j)R^(7k)— or—CR^(7l)R^(7m)—CR^(7n)R^(7o)C(O)—N(R^(7p))—; where each of R^(6a),R^(6b), R^(6c), R^(6d) and R^(6e) is independently selected fromhydrogen and (1-4C)alkyl; and each of R^(7a), R^(7b), R^(7c), R^(7d),R^(7e), R^(7f), R^(7g), R^(7h), R^(7i), R^(7j), R^(7k), R^(7l), R^(7m),R^(7n), R^(7o) and R^(7p) is independently selected from hydrogen and(1-4C)alkyl;

each R^(8a) and R^(8b) is independently selected from hydrogen,(1-4C)alkyl, hydroxy and fluoro, or R^(8a) and R^(8b) are joinedtogether with the atoms to which they are attached to form a(3-6C)cycloalkylene ring or a (2-5C)heterocyclene ring containing 1 or 2heteroatoms independently selected from oxygen, nitrogen and sulfur, orR^(8a) and W^(a) are joined together with the atoms to which they areattached to form a (2-6C)azacycloalkylene group;

a is 0 or an integer of from 1 to 3;

b is an integer of from 2 to 8;

or a pharmaceutically acceptable salt or solvate or stereoisomerthereof.

In a particular aspect, this invention provides a compound of formulaII:

wherein R¹, R², R³, R⁴, R^(8a), R^(8b), W, a and b are as defined herein(including any specific or preferred embodiments); or a pharmaceuticallyacceptable salt or solvate or stereoisomer thereof.

In another particular aspect, this invention provides a compound offormula III:

wherein R¹, R², R³, R⁴, R^(8a), R^(8b), W, a and b are as defined herein(including any specific or preferred embodiments); or a pharmaceuticallyacceptable salt or solvate or stereoisomer thereof.

In yet another particular aspect, this invention provides a compound offormula IV:

wherein R¹, R², R³, R⁴, R^(8a), R^(8b), W, a and b are as defined herein(including any specific or preferred embodiments); or a pharmaceuticallyacceptable salt or solvate or stereoisomer thereof.

In another particular aspect, this invention provides a compound offormula V:

wherein R², R³, R⁴, R⁵, R⁶, R⁷, R^(8a), R^(8b)W, a, and b are as definedherein (including any specific or preferred embodiments); and each R⁹ isindependently selected from (1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl,(3-6C)cycloalkyl, cyano, halo, —OR^(9a), —C(O)OR^(9b), —SR^(9c),—S(O)R^(9d), —S(O)₂R^(9e) and —NR^(9f)R^(9g); wherein R^(9a), R^(9b),R^(9c), R^(9d), R^(9e), R^(9f) and R^(9g) are independently selectedfrom hydrogen, (1-4C)alkyl or phenyl-(1-4C)alkyl; and n is 0, 1, 2 or 3;or a pharmaceutically acceptable salt or solvate or stereoisomerthereof.

In another particular aspect, this invention provides a compound offormula VI:

wherein R², R³, R⁴, R⁵, R⁶, R⁷, R^(8a), R^(8b), W, a and b are asdefined herein (including any specific or preferred embodiments); andeach R¹⁰ is independently selected from (1-4C)alkyl, (2-4C)alkenyl,(2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(10a), —C(O)OR^(10b),—SR^(10c), —S(O)R^(10d), —S(O)₂R^(10e) and —NR^(10f)R^(10g); whereinR^(10a), R^(10b), R^(10c), R^(10d), R^(10e), R^(10f) and R^(10g) areindependently selected from hydrogen, (1-4C)alkyl or phenyl-(1-4C)alkyl;and o is 0, 1, 2 or 3; or a pharmaceutically acceptable salt or solvateor stereoisomer thereof.

In another particular aspect, this invention provides a compound offormula VII:

wherein R², R³, R⁴, R⁵, R⁶, R⁷, R^(8a), R^(8b), W, a and b are asdefined herein (including any specific or preferred embodiments); andeach R¹¹ is independently selected from (1-4C)alkyl, (2-4C)alkenyl,(2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(11a), —C(O)OR^(11b),—SR^(11b), —S(O)R^(11c), —S(O)R^(11d), —S(O)₂R^(11e) and—NR^(11f)R^(11g); wherein R^(11a), R^(11b), R^(11c), R^(11d), R^(11e),R^(11f) and R^(11g) are independently selected from hydrogen,(1-4C)alkyl or phenyl-(1-4C)alkyl; and p is 0, 1 or 2; or apharmaceutically acceptable salt or solvate or stereoisomer thereof.

In another particular aspect, this invention provides a compound offormula VIII:

or the corresponding pyridine N-oxide; wherein R², R³, R⁴, R⁵, R⁶, R⁷,R^(8a), R^(8b), W, a and b are as defined herein (including any specificor preferred embodiments); and each R¹² is independently selected from(1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano,halo, —OR^(12a), —C(O)OR^(12b), —SR^(12c), —S(O)R^(12d), —S(O)₂R^(12e)and NR^(12f)R^(12g); wherein R^(12a), R^(12b), R^(12c), R^(12d),R^(12e), R^(12f) and R^(12g) are independently selected from hydrogen,(1-4C)alkyl or phenyl-(1-4C)alkyl; and q is 0, 1 or 2; or apharmaceutically acceptable salt or solvate or stereoisomer thereof.

In another of its composition aspects, this invention provides apharmaceutical composition comprising a pharmaceutically acceptablecarrier and a therapeutically effective amount of a compound of formulaI or a pharmaceutically acceptable salt or solvate or stereoisomerthereof. Such pharmaceutical compositions may optionally contain othertherapeutic agents. Accordingly, in one embodiment, this invention isdirected to such a pharmaceutical composition wherein the compositionfurther comprises a therapeutically effective amount of a steroidalanti-inflammatory agent, such as a corticosteroid.

Compounds of this invention possess both β₂ adrenergic receptor agonistactivity and muscarinic receptor antagonist activity. Accordingly, thecompounds of formula I are expected to be useful as therapeutic agentsfor treating pulmonary disorders, such as asthma and chronic obstructivepulmonary disease.

Accordingly, in one of its method aspects, this invention provides amethod for treating a pulmonary disorder, the method comprisingadministering to a patient in need of treatment a therapeuticallyeffective amount of a compound of formula I or a pharmaceuticallyacceptable salt or solvate or stereoisomer thereof.

Additionally, in another of its method aspects, this invention providesa method of producing bronchodilation in a patient, the methodcomprising administering to a patient requiring bronchodilation abronchodilation-producing amount of a compound of formula I or apharmaceutically acceptable salt or solvate or stereoisomer thereof.

This invention also provides, a method of treating chronic obstructivepulmonary disease or asthma, the method comprising administering to apatient in need of treatment a therapeutically effective amount of acompound of formula I or a pharmaceutically acceptable salt or solvateor stereoisomer thereof.

Since compounds of this invention possess both β₂ adrenergic receptoragonist activity and muscarinic receptor antagonist activity, suchcompounds are also useful as research tools. Accordingly, in yet anotherof its method aspects, this invention provides a method for using acompound of formula I or a pharmaceutically acceptable salt or solvateor stereoisomer thereof as a research tool for studying a biologicalsystem or sample, or for discovering new chemical compounds having bothβ₂ adrenergic agonist activity and muscarinic receptor antagonistactivity.

This invention also provides processes and novel intermediates usefulfor preparing compounds of formula I or a pharmaceutically acceptablesalt or solvate or stereoisomer thereof. Accordingly, in another of itsmethod aspects, this invention provides a process of preparing acompound of formula I, the process comprising:

(a) reacting a compound of formula 1 or a salt thereof, with a compoundof formula 2;

(b) reacting a compound of formula 3 or a salt thereof, with a compoundof formula 4;

(c) coupling a compound of formula 5 with a compound of formula 6;

(d) for a compound of formula I wherein R⁵ represents a hydrogen atom,reacting a compound of formula 3 with a compound of formula 7 or ahydrate thereof, in the presence of a reducing agent;

(e) reacting a compound of formula I with a compound of formula 8 or ahydrate thereof, in the presence of a reducing agent;

(f) reacting a compound of formula 9, with a compound of formula 10; or

(g) reacting a compound of formula 11 or a hydrate thereof, with acompound of formula 10, in the presence of a reducing agent;

(h) reacting a compound of formula 12, with a compound of the formula:R¹—B(OH)₂ in the presence of a coupling catalyst;

and then removing any protecting groups to form a compound of formula I;wherein the compounds of formula 1-12 are as defined therein.

In one embodiment, the above process further comprises the step offorming a pharmaceutically acceptable salt of a compound of formula I.In other embodiments, this invention is directed to the other processesdescribed herein; and to the product prepared by any of the processesdescribed herein.

In a particular embodiment, this invention is directed to a compound offormula 12, which compound is useful as an intermediate in preparingcompounds of formula I.

This invention provides a compound of formula I or a pharmaceuticallyacceptable salt or solvate or stereoisomer thereof, for use in therapyor as a medicament.

Additionally, this invention is directed to the use of a compound offormula I or a pharmaceutically acceptable salt or solvate orstereoisomer thereof, for the manufacture of a medicament; especiallyfor the manufacture of a medicament for the treatment of a pulmonarydisorder.

DETAILED DESCRIPTION OF THE INVENTION

In one of its composition aspects, this invention is directed to novelcompounds of formula I or pharmaceutically acceptable salts or solvatesor stereoisomers thereof. These compounds contain one or more chiralcenters and therefore, this invention is directed to racemic mixtures;pure stereoisomers (i.e., enantiomers or diastereomers);stereoisomer-enriched mixtures and the like unless otherwise indicated.When a particular stereoisomer is shown or named herein, it will beunderstood by those skilled in the art that minor amounts of otherstereoisomers may be present in the compositions of this inventionunless otherwise indicated, provided that any utility of the compositionas a whole is not eliminated by the presence of such other isomers.

In particular, compounds of formula I contain a chiral center at thecarbon atom indicated by the symbol * in the following formula:

In one embodiment of this invention, the carbon atom identified by thesymbol * has the (R) configuration. In this embodiment, it is preferredfor compounds of formula I to have the (R) configuration at the carbonatom identified by the symbol * or to be enriched in a stereoisomericform having the (R) configuration at this carbon atom. In anotherembodiment of this invention, the carbon atom identified by the symbol *has the (S) configuration. In this embodiment, it is preferred forcompounds of formula I to have the (S) configuration at the carbon atomidentified by the symbol * or to be enriched in a stereoisomeric formhaving the (S) configuration at this carbon atom. In some cases, inorder to optimize the β₂ adrenergic agonist activity of the compounds ofthis invention, it is preferred that the carbon atom identified by thesymbol * has the (R) configuration.

The compounds of formula I also contain several basic groups (e.g.,amino groups) and therefore, the compounds of formula I can exist as thefree base or in various salt forms. All such salt forms are includedwithin the scope of this invention. Furthermore, solvates of compoundsof formula I or salts thereof are included within the scope of thisinvention.

Additionally, where applicable, all cis-trans or E/Z isomers (geometricisomers), tautomeric forms and topoisomeric forms of the compounds offormula I are included within the scope of this invention unlessotherwise specified.

The nomenclature used herein to name the compounds of this invention andintermediates thereof has generally been derived using thecommercially-available AutoNom software (MDL, San Leandro, Calif.).Typically, compounds of formula I wherein W is O have been named asester derivatives of carbamic acid; and compounds of formula I wherein Wis NW^(a) have been named as urea derivatives.

REPRESENTATIVE EMBODIMENTS

The following substituents and values are intended to providerepresentative examples of various aspects and embodiments of thisinvention. These representative values are intended to further defineand illustrate such aspects and embodiments and are not intended toexclude other embodiments or to limit the scope of this invention. Inthis regard, the representation that a particular value or substituentis preferred is not intended in any way to exclude other values orsubstituents from this invention unless specifically indicated.

In one embodiment of this invention, W is O. In another embodiment, W isNW^(a).

Generally, it has been found that compounds in which W represents Oexhibit particularly high affinity for muscarinic and β₂ adrenergicreceptors. Accordingly, in a particular embodiment of this invention, Wpreferably represents O.

When W is NW^(a), W^(a) is hydrogen or (1-4C)alkyl; such as hydrogen,methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl andtert-butyl. In one embodiment, W^(a) is hydrogen or (1-3C)alkyl. Inanother embodiment, W^(a) is hydrogen, methyl or ethyl; such as hydrogenor methyl. In yet another embodiment, W^(a) is hydrogen and NW^(a) isNH.

In a particular embodiment, when W is NW^(a), R⁶ is —N(R^(6a))C(O)R^(6b)(e.g., —NHCHO) and R⁷ is hydrogen; or R⁶ and R⁷ together form—N(R^(7a))C(O)C(R^(7b))═C(R^(7c))—(e.g., —NHC(O)CH═CH—).

In the compounds of the present invention, R¹ is an optionallysubstituted (6-10C)aryl, (2-9C)heteroaryl or (3-7C)cycloalkyl group. Ina particular embodiment, R¹ is an optionally substituted (6-10C)arylgroup selected from a phenyl and naphthalene ring, where the point ofattachment is at any available carbon ring atom. In another particularembodiment, R is an optionally substituted (2-9C)heteroaryl groupselected from a pyrrole, imidazole, thiazole, oxazole, furan, thiophene,triazole, pyrazole, isoxazole, isothiazole, pyridine, pyridine N-oxide,pyrazine, pyridazine, pyrimidine, triazine, indole, benzofuran,benzothiophene, benzimidazole, benzthiazole, quinoline, isoquinoline,quinazoline and quinoxaline ring, where the point of attachment is atany available carbon or nitrogen ring atom. In a particular embodiment,R¹ is an optionally substituted phenyl ring. In another particularembodiment, R¹ is an optionally substituted naphthalene ring, such as anaphth-1-yl or a naphth-2-yl group. In another particular embodiment, R¹is an optionally substituted thienyl group, such as thien-2-yl orthien-3-yl group. In another particular embodiment, R¹ is an optionallysubstituted thiazole group, such as a thiazol-2-yl, thiazol-4-yl orthiazol-5-yl group. In yet another particular embodiment, R¹ is anoptionally substituted pyridyl group, such as a pyrid-2-yl, pyrid-3-ylor pyrid-4-yl group, or the corresponding pyridine N-oxides. In stillanother particular embodiment, R¹ is an optionally substituted furylgroup, such as a fur-2-yl or fur-3-yl group. In another particularembodiment, R¹ is an optionally substituted pyrimidinyl group, such as apyrimidin-2-yl, pyrimidin-4-yl or pyrimidin-4-yl group. In a particularembodiment, R¹ is an optionally substituted cyclopentyl group, such ascyclopentyl, 2,2-difluorocyclopentyl or 3,3-difluorocyclopentyl. Inanother particular embodiment, R¹ is an optionally substitutedcyclohexyl group.

In a particular embodiment, the (6-10C)aryl, (2-9C heteroaryl) or(3-7C)cycloalkyl group of R¹ is unsubstituted or substituted with from 1to 3 substituents selected from (1-4C)alkyl, halo, —OR^(1a) and—NR^(1f)R^(1g), wherein each alkyl group is unsubstituted or substitutedwith from 1 to 3 fluoro substituents. Representative substituentsinclude, but are not limited to, methyl, ethyl, n-propyl, isopropyl,n-butyl, sec-butyl, isobutyl, tert-butyl, fluoro, chloro, bromo, iodo,hydroxy, methoxy, ethoxy, isopropoxy, amino, methylamino, dimethylamino,fluoromethyl, difluoromethyl, trifluoromethyl, 2,2,2,-trifluoroethyl,fluoromethoxy, difluoromethoxy, trifluoromethoxy, 2,2,2-trifluoroethoxyand the like. Particular values for substituents on the R¹ group arefluoro or chloro.

When present, each R² may be at the 3, 4, 5 or 6-position on thephenylene ring to which it is attached (where the carbon atom on thephenylene ring attached to the nitrogen atom is position 1). In oneembodiment, each R² is independently selected from (1-4C)alkyl, halo,—OR^(2a) and —NR^(2f)R^(2g); wherein each alkyl group is unsubstitutedor substituted with from 1 to 3 fluoro substituents. Representativesubstituents include, but are not limited to, methyl, ethyl, n-propyl,isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, fluoro, chloro,bromo, iodo, hydroxy, methoxy, ethoxy, isopropoxy, amino, methylamino,dimethylamino, fluoromethyl, difluoromethyl, trifluoromethyl,2,2,2,-trifluoroethyl, fluoromethoxy, difluoromethoxy, trifluoromethoxy,2,2,2-trifluoroethoxy and the like. Particular values for R² are fluoroor chloro.

In a particular embodiment of the compounds of formula I, a is 0, 1 or2; including 0 or 1. In one embodiment, a is 0.

Each R^(1a), R^(1b), R^(1c), R^(1d), R^(11e), R^(1f), R^(11g), R^(1h),R^(1i), R^(1j), R^(1k), R^(1l), R^(1m),R^(1n),R^(1o), R^(1p) and R^(1q),and R^(2a), R^(2b), R^(2c), R^(2d), R^(2e), R^(2f) and R^(2g) as used inR¹ and R², respectively, is independently hydrogen, (1-4C)alkyl orphenyl-(1-4C)alkyl, where the alkyl group is unsubstituted orsubstituted with from 1 to 3 fluoro substituents. Representative groupsinclude hydrogen, methyl, trifluoromethyl, ethyl, n-propyl, isopropyl,n-butyl, sec-butyl, isobutyl, tert-butyl or benzyl. In one embodiment,these groups are independently hydrogen or (1-3C)alkyl. In anotherembodiment, these groups are independently hydrogen, methyl or ethyl.

In a particular embodiment of the compounds of formula 1, b is 2, 3, 4,5 or 6; including 2, 3, or 4; such as 2 or 3. In one embodiment, b is 2.In another embodiment, b is 3. In yet another embodiment, b is 4.

In one embodiment, each R^(8a) and R^(8b) are independently selectedfrom hydrogen, methyl or ethyl. A particular value for each R^(8a) andR^(8b) is hydrogen or methyl. In a particular embodiment, R^(8a) andR^(8b) in the —CR^(8a)R^(8b) unit attached to W are both methyl, whilein the remaining —CR^(8a)R^(8b) unit or units, R^(8a) and R^(8b) areboth hydrogen. Representative groups in this embodiment include, by wayof illustration, —C(CH₃)₂CH₂—; —C(CH₃)₂CH₂CH₂— and —C(CH₃)₂CH₂CH₂CH₂—.

In another embodiment, R^(8a) and R^(8b) in one —CR^(8a)R^(8b) unit arejoined, together with the carbon atom to which they are attached, toform a (3-6C)cycloalkylene group, such as a cyclopentadi-1,1-yl orcyclohexadi-1,1-yl. Additionally, in another embodiment, R^(8a) andR^(8b) in two —CR^(8a)R^(8b) units are joined, together with the carbonatom to which they are attached, to form a (3-6C)cycloalkylene group,such as a cyclopentadi-1,2-yl, cyclopentadi-1,3-yl, cyclohexadi-1,2-yl,cyclohexadi-1,3-yl or cyclohexadi-1,4-yl group.

In yet another embodiment, R^(8a) in the —CR^(8a) R^(8b) unit attachedto W and W^(a) are joined, together with the atoms to which they areattached, to form a (4-5C)azacycloalkylene group, i.e., apyrrolidin-1,2-yl, pyrrolidin-1,3-yl, piperidin-1,2-yl, piperidin-1,3-ylor piperidin-1,4-yl ring.

In a particular embodiment, R³ is selected from hydrogen or (1-3C)alkyl;such as hydrogen, methyl, ethyl, n-propyl or isopropyl. In anotheraspect, R³ is methyl or ethyl.

In one embodiment, R⁵ is hydrogen or (1-4C)alkyl; such as hydrogen,methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl andtert-butyl. In another embodiment, each R⁵ is independently hydrogen,methyl or ethyl. In a particular embodiment, R⁵ is hydrogen.

In one embodiment, R⁶ is —N(R^(6a))C(O)R^(6b) and R⁷ is hydrogen, whereeach of R^(6a) and R^(6b) is independently hydrogen or (1-4C)alkyl, suchas hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl,isobutyl and tert-butyl. In one embodiment, these groups areindependently hydrogen or (1-3C)alkyl. In another embodiment, thesegroups are independently hydrogen, methyl or ethyl. A particular valuefor R⁶ in this embodiment is —NHCHO.

In another embodiment, R⁶ and R⁷ together form—N(R^(7a))C(O)—CR^(7b)═CR^(7c)—, —CR^(7d)═CR^(7e)—C(O)N(R^(7f))—,—N(R^(7g))C(O)CR^(7h)R^(7i)—CR^(7j)R^(7k)— or—CR^(7l)R^(7m)—CR^(7n)R^(7o)—C(O)N(R^(7p))—; where each of R^(7a),R^(7b), R^(7c), R^(7d), R^(7e), R^(7f), R^(7h), R^(7i), R^(7j), R^(7k),R^(7l), R^(7m), R^(7n), R^(7o) and R^(7p) is independently hydrogen or(1-4C)alkyl; such as hydrogen, methyl, ethyl, n-propyl, isopropyl,n-butyl, sec-butyl, isobutyl and tert-butyl. In one embodiment, thesegroups are independently hydrogen or (1-3C)alkyl. In another embodiment,these groups are independently hydrogen, methyl or ethyl. Particularvalues for R⁶ and R⁷ in this embodiment are R⁶ and R⁷ together form—NHC(O)—CH═CH—, —CH═CH—C(O)—NH—, —CH₂—CH₂—C(O)NH— or —NHC(O)—CH₂—CH₂—;including where R⁶ and R⁷ together form —NHC(O)—CH═CH— or—CH═CH—C(O)—NH—; and in particular, where R⁶ and R⁷ together form—NHC(O)—CH═CH— (i.e., the nitrogen atom is attached at R⁶ and the carbonatom is attached at R⁷ to form, together with the hydroxyphenyl ring towhich R⁶ and R⁷ are attached, a 8-hydroxy-2-oxo-1,2-dihydroquinolin-5-ylgroup).

In a particular embodiment, R⁶ is —NHCHO or —CH₂OH and R⁷ is hydrogen;or R⁶ and R⁷ together form —NHC(O)CH═CH—, —CH═CHC(O)NH—, —CH₂CH₂C(O)NH—or —NHC(O)CH₂CH₂—.

The divalent hydrocarbon group of this invention (e.g., R⁴ in formula 1)contains from 4 to 28 carbon atoms and optionally containing from 1 to10 heteroatoms selected independently from halo, oxygen, nitrogen andsulfur. In one embodiment, this group contains from 4 to 24 carbonatoms, including from 6 to 20 carbon atoms, such as from 8 to 18 carbonatoms; and optionally contains from 1 to 8 heteroatoms, including from 1to 6 heteroatoms.

The divalent hydrocarbon may contain any arrangement of atoms includingalkylene, alkenylene, alkynylene, cycloalkylene, arylene, heteroaryleneand heterocyclene groups or combinations thereof. The hydrocarbon groupmay be interrupted by one or more heteroatoms or combinations ofheteroatoms and carbon atoms to form various functional groups, such asethers, thioethers, amines, amides, esters, carbamates, ureas, sulfones,sulfoxides, sulfonamides and the like.

In a particular aspect of this invention, the divalent hydrocarbon groupof the compounds of this invention (e.g., R⁴ in formula 1) is a divalentgroup of the formula:—(R^(4a))_(d)-(A¹)_(e)-(R^(4b))_(f)-Q-(R^(4c))_(g)-(A²)_(h)-(R^(4d))_(i)—

wherein

d, e, f, g, h and i are each independently selected from 0 and 1;

R^(4a), R^(4b), R^(4c) and R^(4d) are each independently selected from(1-10C)alkylene, (2-10C)alkenylene and (2-10C)alkynylene, wherein eachalkylene, alkenylene or alkynylene group is unsubstituted or substitutedwith from 1 to 5 substituents independently selected from (1-4C)alkyl,fluoro, hydroxy, phenyl and phenyl-(1-4C)alkyl; or R^(4d) represents(1-6C)alkylene-NHC(O)-(1-6C)alkylene;

A¹ and A² are each independently selected from (3-7C)cycloalkylene,(6-10C)arylene, —O-(6-10C)arylene, (6-10C)arylene-O—,(2-9C)heteroarylene, —O-(2-9C)heteroarylene, (2-9C)heteroarylene-O— and(3-6C)heterocyclene, wherein each cycloalkylene is unsubstituted orsubstituted with from 1 to 4 substituents selected independently from(1-4C)alkyl, and each arylene, heteroarylene or heterocyclene group isunsubstituted or substituted with from 1 to 4 substituents independentlyselected from halo, (1-4C)alkyl, (1-4C)alkoxy, —S-(1-4C)alkyl,—S(O)-(1-4C)alkyl, —S(O)₂-(1-4C)alkyl, —C(O)O(1-4C)alkyl, carboxy,cyano, hydroxy, nitro, trifluoromethyl and trifluoromethoxy;

Q is selected from a bond, —O—, —C(O)O—, —OC(O)—, —S—, —S(O)—, —S(O)₂—,—N(Q^(a))C(O)—, —C(O)N(Q^(b))—, —N(Q^(c))S(O)₂—, —S(O)₂N(Q^(d))—,—N(Q^(e))C(O)N(Q^(f))—, —N(Q^(g))S(O)₂N(Q^(h))—, —OC(O)N(Q^(i))—,—N(Q^(j))C(O)O— and —N(Q^(k)); where Q^(a), Q^(b), Q^(c), Q^(d), Q^(e),Q^(f), Q^(g), Q^(h), Q^(i), Q^(j) and Q^(k) are each independentlyselected from hydrogen, (1-6C)alkyl, A³ and (1-4C)alkylene-A⁴, whereinthe alkyl group is unsubstituted or substituted with from 1 to 3substituents independently selected from fluoro, hydroxy and(1-4C)alkoxy; or together with the nitrogen atom and the group R^(4b) orR^(4c) to which they are attached, form a 4 to 6 memberedazacycloalkylene group; and

A³ and A⁴ are each independently selected from (3-6C)cycloalkyl,(6-10C)aryl, (2-9C)heteroaryl and (3-6C)heterocyclyl, wherein eachcycloalkyl is unsubstituted or substituted with from 1 to 4 substituentsselected independently from (1-4C)alkyl and each aryl, heteroaryl orheterocyclyl group is unsubstituted or substituted with from 1 to 4substituents independently selected from halo, (1-4C)alkyl and(1-4C)alkoxy.

In the compound of this invention, the values of each of the componentsR^(4a), A¹, R^(4b), Q, R^(4c), A² and R^(4d) are selected such that thenumber of contiguous atoms in the shortest chain between the twonitrogen atoms to which R⁴ is attached is in the range of from 4 to 16,(specifically, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16);including 8, 9, 10, 11, 12, 13 or 14; such as 8, 9, 10 or 11; or 9 or10. When selecting values for each variable in R⁴, it will beappreciated by those skilled in the art that values should be selectedsuch that a chemically stable group is formed.

When determining the number of contiguous atoms in the shortest chainbetween the two nitrogen atoms to which R⁴ is attached, each contiguousatom of the chain is counted consecutively starting from the first atomin the R⁴ group adjacent to the nitrogen to which R³ is attached andending with the last atom in the R⁴ group adjacent to the nitrogen ofthe aminohydroxyethyl group. Where two or more chains are possible, theshortest chain is used to determine the number of contiguous atoms. Asshown below, for example, when R⁴ is—(CH₂)₂—NHC(O)—CH₂-(phen-1,4-ylene)—CH₂—, there are 10 contiguous atomsin the shortest chain counted consecutively starting from the first atomin the R⁴group adjacent to the nitrogen to which R³ is attached andending with the last atom in the R⁴ group adjacent to the nitrogen ofthe aminohydroxyethyl group as shown below:

In one embodiment of R⁴, R^(4a) is selected from (1-10C)alkylene,(2-10C)alkenylene and (2-10C)alkynylene wherein the alkylene group isunsubstituted or substituted with 1 or 2 substituents independentlyselected from (1-4C)alkyl, hydroxy and phenyl. Representative examplesof particular values for R^(4a) are —CH₂—, —(CH₂)₂—, —(CH₂)₃—, —(CH₂)₄—,—(CH₂)₅—, —(CH₂)₆—, —(CH₂)₇—, —(CH₂)₈—, —(CH₂)₉—, —(CH₂)₁₀—,—(CH₂)CH(CH₃)—, —(CH₂)C(CH₃)₂—, and —(CH₂)₂C(phenyl)₂—. In anotheraspect, R^(4a) is —(CH₂)C(═CH₂)—.

In one embodiment, d is 1.

In one embodiment, A¹ is an optionally substituted (3-7C)cycloalkylenegroup; including a cyclohexylene group, such as cyclohex-1,4-ylene andcyclohex-1,3-ylene; and a cyclopentylene group, such ascyclopent-1,3-ylene.

In another embodiment, A¹ is an optionally substituted (6-10C)arylenegroup, including a phenylene group, such as phen-1,4-ylene,phen-1,3-ylene and phen-1,2-ylene; and a naphthylene group, such asnaphth-1,4-ylene and napth-1,5-ylene.

In yet another embodiment, A¹ is an optionally substituted(2-9C)heteroarylene group, including a pyridylene group, such aspyrid-1,4-ylene; a furylene group, such as fur-2,5-ylene andfur-2,4-ylene; a thienylene group, such as thien-2,5-ylene andthien-2,4-ylene; and a pyrrolylene, such as pyrrol-2,5-ylene andpyrrol-2,4-ylene.

In still another embodiment, A¹ is an optionally substituted(3-6C)heterocyclene group, including a piperidinylene group, such aspiperidin-1,4-ylene; and a pyrrolidinylene group, such aspyrrolidin-2,5-ylene.

In a particular embodiment, A¹ is an optionally substituted phenylene,thienylene, cyclopentylene, cyclohexylene or piperidinylene.

In one embodiment, e is 0.

In a particular embodiment, R^(4b) is (1-5C)alkylene. Representativeexamples of particular values for R^(4b) are —CH₂—, —(CH₂)₂—, —(CH₂)₃—,—(CH₂)₄—, —(CH₂)₅—; including methylene, ethylene and propylene.

In one embodiment, f is 0.

In a particular embodiment, Q is selected from a bond, —O—,—N(Q^(a))C(O)—, —C(O)N(Q^(b))—, —N(Q^(c))S(O)₂—, —S(O)₂N(Q^(d))—,—N(Q^(e))C(O)N(Q^(f))—, —OC(O)N(Q^(i))—, —N(Q^(j))C(O)O— or —N(Q^(k));such as where Q is a bond, —N(Q^(a))C(O)— or —C(O)N(Q^(b)).

Representative examples of particular values for Q are a bond, O, NH,—C(O)NH—, —C(O)N(CH₃)—, —NHC(O)—, —N(CH₃)C(O)—, —S(O)₂NH—,—S(O)₂N(CH₃)—, —NHS(O)₂—, —N(CH₃)S(O)₂— and —NHC(O)NH—. Another exampleof a value for Q, together with R^(4c), is —C(O)(piperidin-1,4-ylene).

In one embodiment, Q^(a), Q^(b), Q^(c), Q^(d), Q^(e), Q^(f), Q^(g),Q^(h), Q^(i), Q^(j) and Q^(k) are each independently selected fromhydrogen and (1-6C)alkyl, wherein the alkyl group is unsubstituted orsubstituted with from 1 to 3 substituents independently selected fromfluoro, hydroxy and (1-4C)alkoxy. For example, Q^(a), Q^(b), Q^(c),Q^(d), Q^(e), Q^(f), Q^(g), Q^(h), Q^(i), Q^(j) and Q^(k) are eachindependently selected from hydrogen, and (1-3C)alkyl, includinghydrogen, methyl, ethyl, n-propyl and isopropyl. An example of a valuefor each of Q^(a), Q^(b), Q^(c), Q^(d), Q^(e), Q^(f), Q^(g), Q^(h),Q^(i), Q^(j) and Q^(k) is hydrogen.

In another embodiment, Q^(a), Q^(b), Q^(c), Q^(d), Q^(e), Q^(f), Q^(g),Q^(h), Q^(i), Q^(j) and Q^(k) together with the nitrogen atom and thegroup R^(4b) or R^(4c) to which they are attached, form a 4-6 memberedazacycloalkylene group. For example, Q^(a) and Q^(b) together with thenitrogen atom and the group R^(4b) or R^(4c) to which they are attached,form a piperidin-4-ylene group. By way of illustration, when Qrepresents —N(Q^(a))C(O)— and Q^(a) together with the nitrogen atom andthe group R^(4b) to which it is attached, forms a piperidin-4-ylenegroup, R⁴ is a group of formula:

Similarly, when Q represents —C(O)N(Q^(b))- and Q^(b) together with thenitrogen atom and the group R^(4c) to which it is attached, forms apiperidin-4-ylene group, R⁴ is a group of formula:

In a particular embodiment, R^(4c) is (1-5C)alkylene. Representativeexamples of particular values for R^(4c) are —CH₂—, —(CH₂)₂—, —(CH₂)₃—,—(CH₂)₄—, —(CH₂)₅—; including methylene, ethylene and propylene.

In one embodiment, A² is an optionally substituted (3-7C)cycloalkylenegroup; including a cyclohexylene group, such as cyclohex-1,4-ylene andcyclohex-1,3-ylene; and a cyclopentylene group, such ascyclopent-1,3-ylene.

In another embodiment, A² is an optionally substituted (6-10C)arylenegroup, including a phenylene group, such as phen-1,4-ylene,phen-1,3-ylene and phen-1,2-ylene; and a naphthylene group, such asnaphth-1,4-ylene and napth-1,5-ylene.

In yet another embodiment, A² is an optionally substituted(2-9C)heteroarylene group, including a pyridylene group, such aspyrid-1,4-ylene; a furylene group, such as fur-2,5-ylene andfur-2,4-ylene; a thienylene group, such as thien-2,5-ylene andthien-2,4-ylene; and a pyrrolylene, such as pyrrol-2,5-ylene andpyrrol-2,4-ylene.

In still another embodiment, A² is an optionally substituted(3-6C)heterocyclene group, including a piperidinylene group, such aspiperidin-1,4-ylene; and a pyrrolidinylene group, such aspyrrolidin-2,5-ylene.

In a particular embodiment, A² is optionally substituted phenylene,thienylene, cyclopentylene, cyclohexylene or piperidinylene.

By way of illustration, either A¹ or A² or both can be phenylene, suchas phen-1,4-ylene or phen-1,3-ylene, where the phenylene group isunsubstituted or substituted with from 1 to 4 substituents independentlyselected from halo, (1-4C)alkyl, (1-4C)alkoxy, —S-(1-4C)alkyl,—S(O)-(1-4C)alkyl, —S(O)₂-(1-4C)alkyl, —C(O)O(1-4C)alkyl, carboxy,cyano, hydroxy, nitro, trifluoromethyl and trifluoromethoxy.Representative examples include phen-1,3-ylene, phen-1,4-ylene,4-chlorophen-1,3-ylene, 6-chlorophen-1,3-ylene, 4-methylphen-1,3-ylene,2-fluorophen-1,4-ylene, 2-chlorophen-1,4-ylene, 2-bromophen-1,4-ylene,2-iodophen-1,4-ylene, 2-methylphen-1,4-ylene, 2-methoxyphen-1,4-ylene,2-trifluoromethoxyphen-1,4-ylene, 3-nitrophen-1,4-ylene,3-chlorophen-1,4-ylene, 2,5-difluorophen-1,4-ylene,2,6-dichlorophen-1,4-ylene, 2,6-diiodophen-1,4-ylene,2-chloro-6-methylphen-1,4-ylene, 2-chloro-5-methoxyphen-1,4-ylene,2,3,5,6-tetrafluorophen-1,4-ylene.

Alternatively, A¹ or A² or both can be cyclopentylene or cyclohexylene;wherein the cyclopentylene or cyclohexylene group is unsubstituted orsubstituted with (1-4C)alkyl. Representative examples includecis-cyclopent-1,3-ylene, trans-cyclopent-1,3-ylene,cis-cyclohex-1,4-ylene and trans-cyclohex-1,4-ylene. A¹ or A² or bothcan also be optionally substituted thienylene or piperidinylene, forexample, thien-2,5-ylene or piperidin-1,4-ylene.

In one embodiment, R^(4d) is selected from (1-10C)alkylene,(2-10C)alkenylene and (2-10C)alkynylene wherein the alkylene isunsubstituted or substituted with 1 or 2 substituents independentlyselected from (1-4C)alkyl, hydroxy and phenyl. Representative examplesof particular values for R^(4d) are —(CH₂)—, —(CH₂)₂—, —(CH₂)₃—,—(CH₂)₄—, —(CH₂)₅—, —(CH₂)₆—, —(CH₂)₇—, —(CH₂)₈—, —(CH₂)₉—, —(CH₂)₁₀—,—(CH₂)CH(CH₃)—, —(CH₂)C(CH₃)₂— and —(CH₂)₂C(phenyl)₂—.

In a particular embodiment, R⁴ is a divalent group of the formula:—(R^(4a))_(d)— where R^(4a) is (4-10C)alkylene. In one aspect of thisembodiment, R⁴ is a divalent group of the formula: —(CH₂)_(j)- where jis 8, 9 or 10. Examples of particular values for R⁴ in this embodimentare —(CH₂)₄—, —(CH₂)₅—, —(CH₂)₆—, —(CH₂)₇—, —(CH₂)₈—, —(CH₂)₉, and—(CH₂)₁₀—; including —(CH₂)₈—, —(CH₂)₉— and —(CH₂)₁₀—.

In another particular embodiment, R⁴ is a divalent group of the formula:—(R^(4a))_(d)-(A²)_(h)-(R^(4d))_(i)—

where R^(4a) is (1-10C)alkylene, such as —(CH₂)—, —(CH₂)₂—, —(CH₂)₃—; A²is (6-10C)arylene, such as phen-1,4-ylene or phen-1,3-ylene, or(2-9C)heteroarylene, such as thien-2,5-ylene or thien-2,4-ylene; andR^(4d) is (1-10C)alkylene, such as —(CH₂)—, —(CH₂)₂—, —(CH₂)₃—. Examplesof particular values for R⁴ in this embodiment are—(CH₂)-(phen-1,4-ylene)-(CH₂)—; —(CH₂)-(phen-1,4-ylene)—(CH₂)₂—;—(CH₂)-(phen-1,4-ylene)-(CH₂)₃—; —(CH₂)₂-(phen-1,4-ylene)-(CH₂)—;—(CH₂)₂-(phen-1,4-ylene)—(CH₂)₂—; (CH₂)₂-(phen-1,4-ylene)-(CH₂)₃—;—(CH₂)₃-(phen-1,4-ylene)-(CH₂)—; —(CH₂)₃-(phen-1,4-ylene)-(CH₂)₂—,—(CH₂)₃-(phen-1,4-ylene)-(CH₂)₃—, CH₂)₄-(phen-1,4-ylene)-(CH₂)—;—(CH₂)₄-(phen-1,4-ylene)-(CH₂)₂— and —(CH₂)₄-(phen-1,4-ylene)-(CH₂)₃—.

In yet another particular embodiment, R⁴ is a divalent group of theformula:—(R^(4a))_(d)-Q-(A²)_(h)-(R^(4d))_(i)—where Q is —O— or —N(Q^(k))—; Q^(k) is hydrogen or (1-3C)alkyl, such asmethyl or ethyl; R^(4a) is (1-10C)alkylene, such as —(CH₂)—, —(CH₂)₂—,—(CH₂)₃—; A² is (6-10C)arylene, such as phen-1,4-ylene orphen-1,3-ylene, or (2-9C)heteroarylene, such as thien-2,5-ylene orthien-2,4-ylene; and R^(4d) is (1-10C)alkylene, such as —(CH₂)—,—(CH₂)₂—, —(CH₂)₃—. In this embodiment, the (6-10C)arylene or(2-9C)heteroarylene group is unsubstituted or substituted with from 1 to3 substituents independently selected from halo, (1-4C)alkyl,(1-4C)alkoxy, —S-(1-4C)alkyl, —S(O)-(1-4C)alkyl, —S(O)₂-(1-4C)alkyl,—C(O)O(1-4C)alkyl, carboxy, cyano, hydroxy, nitro, trifluoromethyl andtrifluoromethoxy; such as methyl, ethyl, n-propyl, isopropyl, n-butyl,sec-butyl, isobutyl, tert-butyl, fluoro, chloro, bromo, iodo, hydroxy,methoxy, ethoxy, isopropoxy, trifluoromethyl and trifluoromethoxy, andthe like. Examples of particular values for R⁴ in this embodiment areCH₂)₂—O-(phen-1,4-ylene)-(CH₂)—; —(CH₂)₂—O-(phen-1,4-ylene)-(CH₂)₂—;—(CH₂)₂—O-(phen-1,4-ylene)-(CH₂)₃—; —(CH₂)₃—O-(phen-1,4-ylene)-(CH₂)—;—(CH₂)₃—O-(phen-1,4-ylene)-CH₂)₂—; —(CH₂)₃—O-(phen-1,4-ylene)-(CH₂)₃—;—(CH₂)₂—NH-(phen-1,4-ylene)-(CH₂)—; —(CH₂)₂—NH-(phen-1,4-ylene)-(CH₂)₂—;—(CH₂)₂—NH-(phen-1,4-ylene)-(CH₂)₃—; CH₂)₃—NH-(phen-1,4-ylene)-(CH₂)—;—(CH₂)₃—NH-(phen-1,4-ylene)-(CH₂)₂—;—(CH₂)₃—NH-(phen-1,4-ylene)-(CH₂)₃—;—(CH₂)₂—NH-(2-fluorophen-1,4-ylene)-(CH₂)—;—(CH₂)₂—NH-(2-chlorophen-1,4-ylene)-(CH₂)—;—(CH₂)₂—NH-(5-methoxyphen-1,4-ylene)-(CH₂)—;—(CH₂)₂—NH-(2-chloro-5-methoxyphen-1,4-ylene)-(CH₂)—; and—(CH₂)₂—NH-(2-chloro-5-ethoxyphen-1,4-ylene)-(CH₂)—.

In yet another particular embodiment, R⁴ is a divalent group of theformula:—(R^(4a))_(d)-(A¹)_(e)-(R^(4b))_(f)-Q-(R^(4c))_(g)-(A²)_(h)-(R^(4d))_(i)—

where Q is —N(Q^(a))C(O) or —C(O)N(Q^(b)). A particular value for R⁴ inthis embodiment is the formula:

where za is an integer from 2 to 10; and zb is an integer from 2 to 10;provided that za+zb is an integer from 4 to 12. In this formula for R⁴,d and g are 1 and e, f, h and i are 0; and R^(4a) is —(CH₂)_(za)—,R^(4c) is —(CH₂)_(zb)— and Q is —C(O)NH—. Particular values for za are 2or 3; and for zb, 4, 5 or 6.

Another particular value for R⁴ is the formula:

where zc is an integer from 2 to 7; and zd is an integer from 1 to 6;provided that zc+zd is an integer from 3 to 8. In this formula for R⁴,d, h and i are 1 and e, f and g are 0; and R^(4a) is —(CH₂)_(zc)—, A² isphen-1,4-ylene, R^(4d) is —(CH₂)_(zd)— and Q is —C(O)NH—. Particularvalues for zc are 2 or 3; and for zd, 1 or 2. In this embodiment, thephen-1,4-ylene group may be optionally substituted as defined herein forA².

Another particular value for R⁴ is the formula:

where ze is an integer from 2 to 6; zf is an integer from 1 to 5; and zgis an integer from 1 to 5; provided that ze+zf+zg is an integer from 4to 8. In this formula for R⁴, d, g, h and i are 1 and e and f are 0; andR^(4a) is —(CH₂)_(ze)—, R^(4c) is —(CH₂)_(zf)—, A² is 1,4-phenylene,R^(4d) is —(CH₂)_(zg)— and Q is —C(O)NH—. Particular values for ze are 2or 3; for zf, 1 or 2; and for zg, 1 or 2. In this embodiment, thephen-1,4-ylene group may be optionally substituted as defined herein forA².

Another particular value for R⁴ is the formula:

where zh is an integer from 2 to 10; and zi is an integer from 2 to 10;provided that zh+zi is an integer from 4 to 12. In this formula for R⁴,d and g are 1 and e, f, h and i are 0; and R^(4a) is —(CH₂)_(zh)—,R^(4c) is —(CH₂)_(zi)— and Q is —NHC(O)—. Particular values for zh are 2or 3; and for zi, 4, 5 or 6.

Another particular value for R⁴ is the formula:

where zj is an integer from 2 to 7; and zk is an integer from 1 to 6;provided that zj+zk is an integer from 3 to 8. In this formula for R⁴,d, h and i are 1 and e, f and g are 0; and R^(4a) is —(CH₂)_(zj)—, A² is1,4-phenylene, R^(4d) is CH₂)_(zk)— and Q is —NHC(O)—. Particular valuesfor zj are 2 or 3; and for zk, 1 or 2. In this embodiment, thephen-1,4-ylene group may be optionally substituted as defined herein forA².

Another particular value for R⁴ is the formula:

where zl is an integer from 2 to 6; zm is an integer from 1 to 5; and znis an integer from 1 to 5; provided that zi+zm+zn is an integer from 4to 8. In this formula for R⁴, d, g, h and i are 1 and e and f are 0; andR^(4a) is —(CH₂)_(zi)—, R^(4c) is —(CH₂)_(zm)—, A² is 1,4-phenylene,R^(4d) is —(CH₂)_(zn)— and Q is —NHC(O)—. Particular values for zl are 2or 3; for zm, 1 or 2; and

for zn, 1 or 2. In this embodiment, the phen-1,4-ylene group may beoptionally substituted as defined herein for A².

By way of further illustration, R⁴ can be selected from:

-   —(CH₂)₇—;-   —(CH₂)₈—;-   —(CH₂)₉—;-   —(CH₂)₁₀—;-   —(CH₂)₁₁—;-   —(CH₂)₂C(O)NH(CH₂)₅—;-   —(CH₂)₂N(CH₃)C(O)(CH₂)₅—;-   —(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂—;-   —(CH₂)₂NHC(O)(phen-1,4-ylene)CH₂—;-   —(CH₂)₂NHC(O)NH(CH₂)₅—;-   —(CH₂)₃NHC(O)NH(CH₂)₅—;-   —(CH₂)₂C(O)NHCH₂(cyclohex-1,3-ylene)CH₂—;-   —(CH₂)₂NHC(O)(cyclopent-1,3-ylene)-;-   —(CH₂)₂NHC(O)NH(phen-1,4-ylene)(CH₂)₂—;-   1-[-(CH₂)₂C(O)](piperidin-4-yl)(CH₂)₂—;-   —(CH₂)₂NHC(O)(trans-cyclohex-1,4-ylene)CH₂—;-   —(CH₂)₂NHC(O)(cis-cyclopent-1,3-ylene)-;-   —(CH₂)₂NH(phen-1,4-ylene)(CH₂)₂—;-   1-[-(CH₂)₂NHC(O)](piperidin-4-yl)(CH₂)₂—;-   —CH₂(phen-1,4-ylene)NH(phen-1,4-ylene)CH₂—;-   —(CH₂)₂C(O)NHCH₂(phen-1,3-ylene)CH₂—;-   —(CH₂)₂C(O)NHCH₂(pyrid-2,6-ylene)CH₂—;-   —(CH₂)₂C(O)NH(cis-cyclohex-1,4-ylene)CH₂—;-   —(CH₂)₂C(O)NH(trans-cyclohex-1,4-ylene)CH₂—;-   —(CH₂)₂NHC(O)(cis-cyclopent-1,3-ylene)CH₂—;-   —(CH₂)₂N(CH₃)C(O)(phen-1,3-ylene)CH₂—;-   —(CH₂)₂N(CH₃)C(O)(trans-cyclohex-1,4-ylene)CH₂—;-   —(CH₂)₂C(O)NH(phen-1,4-ylene)C*H(CH₃)—((S)-isomer);-   —(CH₂)₂C(O)NH(phen-1,4-ylene)C*H(CH₃)—((R)-isomer);-   2-[(S)—(—CH₂-](pyrrolidin-1-yl)C(O)(CH₂)₄—;-   2-[(S)—(—CH₂-](pyrrolidin-1-yl)C(O)(phen-1,4-ylene)CH₂—;-   —(CH₂)₂C(O)NH(4-chlorophen-1,3-ylene)CH₂—;-   —CH₂(2-fluorophen-1,3-ylene)CH₂—;-   —(CH₂)₂C(O)NH(4-methylphen-1,3-ylene)CH₂—;-   —(CH₂)₂C(O)NH(6-chlorophen-1,3-ylene)CH₂—;-   —(CH₂)₂C(O)NH(2-chlorophen-1,4-ylene)CH₂—;-   —(CH₂)₂C(O)NH(2,6-dichlorophen-1,4-ylene)CH₂—;-   —(CH₂)₂NHC(O)NHCH₂(phen-1,3-ylene)CH₂—;-   4-[-CH₂-](piperidin-1-yl)C(O)(phen-1,4-ylene)CH₂—;-   —(CH₂)₂C(O)N(CH₂CH₃)(phen-1,4-ylene)CH₂—;-   1-[-(CH₂)₂NHC(O)](piperidin-4-yl)-;-   —(CH₂)₂C(O)NH(phen-1,4-ylene)(CH₂)₂—;-   —(CH₂)₂NHC(O)(thien-2,5-ylene)CH₂—;-   —(CH₂)₂N(CH₃)C(O)(3-nitrophen-1,4-ylene)CH₂—;-   —(CH₂)₂N(CH₃)C(O)(trans-cyclohex-1,4-ylene)-;-   1-[-CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)-;-   5-[-(CH₂)₂NHC(O)](pyrid-2-yl)CH₂—;-   —(CH₂)₂(phen-1,4-ylene)(CH₂)₂—;-   —(CH₂)₃(thien-2,5-ylene)(CH₂)₃—;-   —(CH₂)₂(phen-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂—;-   —CH₂(phen-1,2-ylene)NH(phen-1,4-ylene)(CH₂)₂—;-   1-[-CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)(CH₂)₂—;-   1-[-CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)CH₂—;-   —(CH₂)₂C(O)NH(3-chlorophen-1,4-ylene)CH₂—;-   —(CH₂)₂C(O)NH(2-(CF₃O—)phen-1,4-ylene)CH₂—;-   —(CH₂)₃(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂—;-   —(CH₂)₂S(O)₂NH(CH₂)₅—;-   —CH₂(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂—;-   —(CH₂)₂C(O)NH(2-iodophen-1,4-ylene)CH₂—;-   —(CH₂)₂C(O)NH(2-chloro-5-methoxyphen-1,4-ylene)CH₂—;-   —(CH₂)₂C(O)NH(2-chloro-6-methylphen-1,4-ylene)CH₂—;-   —(CH₂)₂N(CH₃)S(O)₂(phen-1,4-ylene)CH₂—;-   —(CH₂)₂C(O)NH(2-bromophen-1,4-ylene)CH₂—;-   —(CH₂)₃(phen-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂—;-   —(CH₂)₃(phen-1,2-ylene)NH(phen-1,4-ylene)(CH₂)₂—;-   1-[-CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)(CH₂)₃—;-   —(CH₂)₂C(O)NH(2-methoxyphen-1,4-ylene)CH₂—;-   —(CH₂)₅NH(phen-1,4-ylene)(CH₂)₂—;-   4-[-(CH₂)₂-](piperidin-1-yl)(phen-1,4-ylene)(CH₂)₂—;-   —(CH₂)₂C(O)NH(phen-1,4-ylene)CH(CH₃)CH₂—;-   —(CH₂)₂-(trans-cyclohex-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂—;-   —(CH₂)₂C(O)NH(2-fluorophen-1,4-ylene)CH₂—;-   —(CH₂)₂(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂—;-   —(CH₂)₂C(O)NH(2,5-difluorophen-1,4-ylene)CH₂—;-   —(CH₂)₂NHC(O)(phen-1,4-ylene)(CH₂)₂—;-   1-[-CH₂(pyrid-2,6-ylene)CH₂](piperidin-4-yl)CH₂—;-   —(CH₂)₃NH(phen-1,4-ylene)(CH₂)₂—;-   —(CH₂)₂NH(naphth-1,4-ylene)(CH₂)₂—;-   —(CH₂)₃O(phen-1,4-ylene)CH₂—;-   1-[-(CH₂)₃](piperidin-4-yl))CH₂—;-   4-[-(CH₂)₂](piperidin-1-yl)C(O)(phen-1,4-ylene)CH₂—;-   —(CH₂)₃(phen-1,4-ylene)NHC(O)(CH₂)₂—;-   —(CH₂)₃O(phen-1,4-ylene)(CH₂)₂—;-   2-[-(CH₂)₂](benzimidazol-5-yl)CH₂—;-   —(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₂—;-   —(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₄—;-   —(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₅—;-   4-[-(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₂—;-   —(CH₂)₂NHC(O)NH(phen-1,4-ylene)CH₂—;-   —(CH₂)₂N(CH₃)(CH₂)₂(cis-cyclohex-1,4-ylene)-;-   —(CH₂)₂C(O)NH(2,3,5,6-tetrafluorophen-1,4-ylene)CH₂—;-   —(CH₂)₂C(O)NH(2,6-diiodophen-1,4-ylene)CH₂—;-   4-[-(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₃—;-   4-[-(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₄—;-   4-[-(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₅—;-   —(CH₂)₂C(O)NHCH₂(phen-1,4-ylene)CH₂—;-   —(CH₂)₂NHC(O)NHCH₂(phen-1,4-ylene)CH₂—;-   —(CH₂)₂C(O)NH(2-methylphen-1,4-ylene)CH₂—;-   1-[-(CH₂)₃O(phen-1,4-ylene)(CH₂)₂](piperidin-4-yl)CH₂—;-   —(CH₂)₂C(O)NHCH₂(phen-1,3-ylene)(CH₂)₂—;-   —(CH₂)₂O(phen-1,3-ylene)CH₂—;-   —(CH₂)₂N(CH₃)C(O)CH₂O(phen-1,4-ylene)CH₂—;-   —(CH₂)₂N(CH₃)C(O)CH₂O(phen-1,3-ylene)CH₂—;-   —(CH₂)₂N(CH₃)C(O)(fur-2,5-ylene)CH₂—;-   —(CH₂)₂N(CH₃)C(O)(thien-2,5-ylene)CH₂—;-   —(CH₂)₂O(phen-1,4-ylene)O(CH₂)₂—;-   —(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(phen-1,4-ylene)CH₂—;-   —(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)CH₂O(phen-1,2-ylene)CH₂—;-   —(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)CH₂O(phen-1,3-ylene)CH₂—;-   —(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)CH₂O(phen-1,4-ylene)CH₂—;-   —(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(fur-2,5-ylene)CH₂—;-   —(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(thien-2,5-ylene)CH₂—;-   4-[-(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,2-ylene)CH₂—;-   4-[-(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,3-ylene)CH₂—;-   4-[-(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,4-ylene)CH₂—;-   4-[-(CH₂)₂](piperidin-1-yl)C(O)(fur-2,5-ylene)CH₂—;-   4-[-(CH₂)₂](piperidin-1-yl)C(O)(thien-2,5-ylene)CH₂—;-   —(CH₂)₂(phen-1,4-ylene)NHC(O)(phen-1,3-ylene)CH₂—;-   —(CH₂)₂(phen-1,4-ylene)NHC(O)(phen-1,4-ylene)CH₂—;-   —(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,2-ylene)CH₂—;-   —(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,3-ylene)CH₂—;-   —(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,4-ylene)CH₂—;-   —(CH₂)₂(phen-1,4-ylene)NHC(O)(fur-2,5-ylene)CH₂—;-   —(CH₂)₂(phen-1,4-ylene)NHC(O)(thien-2,5-ylene)CH₂—;-   —(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(phen-1,3-ylene)CH₂—;-   —(CH₂)₃O(phen-1,3-ylene)CH₂—;-   —CH₂CH(OH)CH₂NH(phen-1,4-ylene)(CH₂)₂—;-   —(CH₂)₄NH(phen-1,4-ylene)(CH₂)₂—;-   —(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂NHC(O)CH₂—;-   —(CH₂)₂C(O)NH(phen-1,4-ylene)(CH₂)₂NHC(O)CH₂—;-   —(CH₂)₂C(O)NHCH₂(trans-cyclohex-1,4-ylene)CH₂—;-   —(CH₂)₂NHC(O)(CH₂)₅—;-   —(CH₂)₂O(phen-1,3-ylene)O(CH₂)₂—;-   —(CH₂)₂O(phen-1,2-ylene)O(CH₂)₂—;-   —CH₂(phen-1,2-ylene)O(phen-1,2-ylene)CH₂—;-   —(CH₂)₂C(O)NH(CH₂)₆—;-   —(CH₂)₃(phen-1,4-ylene)(CH₂)₃—;-   —(CH₂)₃(phen-1,4-ylene)(CH₂)₂—;-   —(CH₂)₄(phen-1,4-ylene)(CH₂)₂—;-   —(CH₂)₃(furan-2,5-ylene)(CH₂)₃—;-   —(CH₂)₂N(CH₃)C(O)NH(phen-1,4-ylene)(CH₂)₂—;-   4-[-(CH₂)₂](piperidin-1-yl)C(O)NH(phen-1,4-ylene)(CH₂)₂—;-   —(CH₂)₃(phen-1,3-ylene)(CH₂)₃—;-   —(CH₂)₃(tetrahydrofuran-2,5-ylene)(CH₂)₃—; and-   —(CH₂)₂O(phen-1,4-ylene)C(O)(CH₂)₂—.    Representative Subgeneric Groupings

The following subgeneric formulae and groupings are intended to providerepresentative examples of various aspects and embodiments of thisinvention and as such, they are not intended to exclude otherembodiments or to limit the scope of this invention unless otherwiseindicated.

A particular group of compounds of formula I are those where: a is 0; bis 2, 3 or 4; W is O; R¹ is phenyl; R⁵ is hydrogen; and R³, R⁴, R⁶, R⁷,R^(8a) and R^(8b) are as defined herein; or a pharmaceuticallyacceptable salt or solvate or stereoisomer thereof.

Yet another particular group of compounds of formula I are thosewherein: a is 0; b is 2, 3 or 4; W is 0; R¹ is phenyl; R⁴ is —(CH₂)_(j)—where j is 8, 9 or 10; R⁵ is hydrogen; and R³, R⁴, R⁶, R⁷, R^(8a) andR^(8b) are as defined herein; or a pharmaceutically acceptable salt orsolvate or stereoisomer thereof.

Another particular group of compounds of formula I are those of formulaII as defined herein; or a pharmaceutically acceptable salt or solvateor stereoisomer thereof.

Another particular group of compounds of formula I are those of formulaIII as defined herein; or a pharmaceutically acceptable salt or solvateor stereoisomer thereof.

Another particular group of compounds of formula I are those of formulaIV as defined herein; or a pharmaceutically acceptable salt or solvateor stereoisomer thereof.

Another particular group of compounds of formula I are those of formulaV as defined herein; or a pharmaceutically acceptable salt or solvate orstereoisomer thereof.

Another particular group of compounds of formula I are those of formulaVI as defined herein; or a pharmaceutically acceptable salt or solvateor stereoisomer thereof.

Another particular group of compounds of formula I are those of formulaVII as defined herein; or a pharmaceutically acceptable salt or solvateor stereoisomer thereof.

Another particular group of compounds of formula I are those of formulaVIII as defined herein; or a pharmaceutically acceptable salt or solvateor stereoisomer thereof.

Another particular group of compounds of formula I are those selectedfrom:

biphenyl-2-ylcarbamic acid3-({9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydro-quinolin-5-yl)ethylamino]nonyl}methylamino)-1,1-dimethylpropylester;

(2-thien-3-ylphenyl)carbamic acid3-({9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}methylamino)-1,1-dimethylpropylester;

biphenyl-2-ylcarbamic acid3-{[2-(4-{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzoylamino)ethyl]methylamino}-1,1-dimethylpropylester;

biphenyl-2-ylcarbamic acid3-{[4-(4-{2-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenyl)butyl]methylamino}-1,1-dimethyl-propylester; and

biphenyl-2-ylcarbamic acid3-{[2-(2-chloro-4-{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-5-methoxyphenylcarbamoyl)ethyl]-methylamino}-1,1-dimethylpropylester.

Definitions

When describing the compounds, compositions, methods and processes ofthis invention, the following terms have the following meanings unlessotherwise indicated.

The term “alkyl” means a monovalent saturated hydrocarbon group whichmay be linear or branched. Unless otherwise defined, such alkyl groupstypically contain from 1 to 10 carbon atoms. Representative alkyl groupsinclude, by way of example, methyl, ethyl, n-propyl, isopropyl, n-butyl,sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl,n-nonyl, n-decyl and the like.

The term “alkylene” means a divalent saturated hydrocarbon group whichmay be linear or branched. Unless otherwise defined, such alkylenegroups typically contain from 1 to 10 carbon atoms. Representativealkylene groups include, by way of example, methylene, ethane-1,2-diyl(“ethylene”), propane-1,2-diyl, propane-1,3-diyl, butane-1,4-diyl,pentane-1,5-diyl and the like.

The term “alkoxy” means a monovalent group of the formula (alkyl)-O—,where alkyl is as defined herein. Representative alkoxy groups include,by way of example, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy,sec-butoxy, isobutoxy, tert-butoxy and the like.

The term “alkenyl” means a monovalent unsaturated hydrocarbon groupwhich may be linear or branched and which has at least one, andtypically 1, 2 or 3, carbon-carbon double bonds. Unless otherwisedefined, such alkenyl groups typically contain from 2 to 10 carbonatoms. Representative alkenyl groups include, by way of example,ethenyl, n-propenyl, isopropenyl, n-but-2-enyl, n-hex-3-enyl and thelike. The term “alkenylene” means a divalent alkenyl group.

The term “alkynyl” means a monovalent unsaturated hydrocarbon groupwhich may be linear or branched and which has at least one, andtypically 1, 2 or 3, carbon-carbon triple bonds. Unless otherwisedefined, such alkynyl groups typically contain from 2 to 10 carbonatoms. Representative alkynyl groups include, by way of example,ethynyl, n-propynyl, n-but-2-ynyl, n-hex-3-ynyl and the like. The term“alkynylene” means a divalent alkynyl group.

The term “aryl” means a monovalent aromatic hydrocarbon having a singlering (i.e., phenyl) or fused rings (i.e., naphthalene). Unless otherwisedefined, such aryl groups typically contain from 6 to 10 carbon ringatoms. Representative aryl groups include, by way of example, phenyl andnaphthalene-1-yl, naphthalene-2-yl, and the like. The term “arylene”means a divalent aryl group.

The term “azacycloalkyl” means a monovalent heterocyclic ring containingone nitrogen atom, i.e., a cycloalkyl group in which one carbon atom hasbeen replaced with a nitrogen atom. Unless otherwise defined, suchazacycloalkyl groups typically contain from 2 to 9 carbon atoms.Representative examples of an azacycloalkyl group are pyrrolidinyl andpiperidinyl groups. The term “azacycloalkylene” means a divalentazacycloakyl group. Representative examples of an azacycloalkylene groupare pyrrolidinylene and piperidinylene groups.

The term “cycloalkyl” means a monovalent saturated carbocyclichydrocarbon group. Unless otherwise defined, such cycloalkyl groupstypically contain from 3 to 10 carbon atoms. Representative cycloalkylgroups include, by way of example, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl and the like. The term “cycloalkylene” means a divalentcycloalkyl group.

The term “halo” means fluoro, chloro, bromo and iodo.

The term “heteroaryl” means a monovalent aromatic group having a singlering or two fused rings and containing in the ring at least oneheteroatom (typically 1 to 3 heteroatoms) selected from nitrogen, oxygenor sulfur. Unless otherwise defined, such heteroaryl groups typicallycontain from 5 to 10 total ring atoms. Representative heteroaryl groupsinclude, by way of example, monovalent species of pyrrole, imidazole,thiazole, oxazole, furan, thiophene, triazole, pyrazole, isoxazole,isothiazole, pyridine, pyrazine, pyridazine, pyrimidine, triazine,indole, benzofuran, benzothiophene, benzimidazole, benzthiazole,quinoline, isoquinoline, quinazoline, quinoxaline and the like, wherethe point of attachment is at any available carbon or nitrogen ringatom. The term “heteroarylene” means a divalent heteroaryl group.

The term “heterocyclyl” or “heterocyclic” means a monovalent saturatedor unsaturated (non-aromatic) group having a single ring or multiplecondensed rings and containing in the ring at least one heteroatom(typically 1 to 3 heteroatoms) selected from nitrogen, oxygen or sulfur.Unless otherwise defined, such heterocyclic groups typically containfrom 2 to 9 total ring carbon atoms. Representative heterocyclic groupsinclude, by way of example, monovalent species of pyrrolidine,imidazolidine, pyrazolidine, piperidine, 1,4-dioxane, morpholine,thiomorpholine, piperazine, 3-pyrroline and the like, where the point ofattachment is at any available carbon or nitrogen ring atom. The term“heterocyclene” means a divalent heterocyclyl or heterocyclic group.

When a specific number of carbon atoms is intended for a particular termused herein, the number of carbon atoms is shown in parenthesespreceding the term. For example, the term “(1-4C)alkyl” means an alkylgroup having from 1 to 4 carbon atoms.

The term “pharmaceutically acceptable salt” means a salt which isacceptable for administration to a patient, such as a mammal (e.g.,salts having acceptable mammalian safety for a given dosage regime).Such salts can be derived from pharmaceutically acceptable inorganic ororganic bases and from pharmaceutically acceptable inorganic or organicacids. Salts derived from pharmaceutically acceptable inorganic basesinclude ammonium, calcium, copper, ferric, ferrous, lithium, magnesium,manganic, manganous, potassium, sodium, zinc and the like. Salts derivedfrom pharmaceutically acceptable organic bases include salts of primary,secondary and tertiary amines, including substituted amines, cyclicamines, naturally-occurring amines and the like, such as arginine,betaine, caffeine, choline, N,N′-dibenzylethylenediamine, diethylamine,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperadine, polyamine resins,procaine, purines, theobromine, triethyl amine, trimethylamine,tripropyl amine, tromethamine and the like. Salts derived frompharmaceutically acceptable acids include acetic, ascorbic,benzenesulfonic, benzoic, camphosulfonic, citric, ethanesulfonic,edisylic, fumaric, gentisic, gluconic, glucoronic, glutamic, hippuric,hydrobromic, hydrochloric, isethionic, lactic, lactobionic, maleic,malic, mandelic, methanesulfonic, mucic, naphthalenesulfonic,naphthalene-1,5-disulfonic, naphthalene-2,6-disulfonic, nicotinic,nitric, orotic, pamoic, pantothenic, phosphoric, succinic, sulfuric,tartaric, p-toluenesulfonic, xinafoic and the like.

The term “salt thereof” means a compound formed when the hydrogen of anacid is replaced by a cation, such as a metal cation or an organiccation and the like. Preferably, the salt is a pharmaceuticallyacceptable salt, although this is not required for salts of intermediatecompounds that are not intended for administration to a patient.

The term “solvate” means a complex or aggregate formed by one or moremolecules of a solute, i.e. a compound of formula I or apharmaceutically acceptable salt thereof, and one or more molecules of asolvent. Such solvates are typically crystalline solids having asubstantially fixed molar ratio of solute and solvent. Representativesolvents include, by way of example, water, methanol, ethanol,isopropanol, acetic acid and the like. When the solvent is water, thesolvate formed is a hydrate.

It will be appreciated that the term “or a pharmaceutically acceptablesalt or solvate or stereoisomer thereof” is intended to include allpermutations of salts, solvates and stereoisomers, such as a solvate ofa pharmaceutically acceptable salt of a stereoisomer of a compound offormula I.

The term “therapeutically effective amount” means an amount sufficientto effect treatment when administered to a patient in need of treatment.

The term “treating” or “treatment” as used herein means the treating ortreatment of a disease or medical condition (such as COPD) in a patient,such as a mammal (particularly a human) that includes:

-   -   (a) preventing the disease or medical condition from occurring,        i.e., prophylactic treatment of a patient;    -   (b) ameliorating the disease or medical condition, i.e.,        eliminating or causing regression of the disease or medical        condition in a patient;    -   (c) suppressing the disease or medical condition, i.e., slowing        or arresting the development of the disease or medical condition        in a patient; or    -   (d) alleviating the symptoms of the disease or medical condition        in a patient.

The term “leaving group” means a functional group or atom which can bedisplaced by another functional group or atom in a substitutionreaction, such as a nucleophilic substitution reaction. By way ofexample, representative leaving groups include chloro, bromo and iodogroups; sulfonic ester groups, such as mesylate, tosylate, brosylate,nosylate and the like; and acyloxy groups, such as acetoxy,trifluoroacetoxy and the like.

The term “protected derivatives thereof” means a derivative of thespecified compound in which one or more functional groups of thecompound are protected from undesired reactions with a protecting orblocking group. Functional groups which may be protected include, by wayof example, carboxylic acid groups, amino groups, hydroxyl groups, thiolgroups, carbonyl groups and the like. Representative protecting groupsfor carboxylic acids include esters (such as a p-methoxybenzyl ester),amides and hydrazides; for amino groups, carbamates (such astert-butoxycarbonyl) and amides; for hydroxyl groups, ethers and esters;for thiol groups, thioethers and thioesters; for carbonyl groups,acetals and ketals; and the like. Such protecting groups are well-knownto those skilled in the art and are described, for example, in T. W.Greene and G. M. Wuts, Protecting Groups in Organic Synthesis, ThirdEdition, Wiley, New York, 1999, and references cited therein.

The term “amino-protecting group” means a protecting group suitable forpreventing undesired reactions at an amino group. Representativeamino-protecting groups include, but are not limited to,tert-butoxycarbonyl (BOC), trityl (Tr), benzyloxycarbonyl (Cbz),9-fluorenylmethoxycarbonyl (Fmoc), formyl, trimethylsilyl (TMS),tert-butyldimethylsilyl (TBS), and the like.

The term “carboxy-protecting group” means a protecting group suitablefor preventing undesired reactions at a carboxy group. Representativecarboxy-protecting groups include, but are not limited to, esters, suchas methyl, ethyl, tert-butyl, benzyl (Bn), p-methoxybenzyl (PMB),9-fluorenylmethyl (Fm), trimethylsilyl (TMS), tert-butyldimethylsilyl(TBS), diphenylmethyl (benzhydryl, DPM) and the like.

The term “hydroxyl-protecting group” means a protecting group suitablefor preventing undesirable reactions at a hydroxyl group. Representativehydroxyl-protecting groups include, but are not limited to, silyl groupsincluding tri(1-6C)alkylsilyl groups, such as trimethylsilyl (TMS),triethylsilyl (TES), tert-butyldimethylsilyl (TBS) and the like; esters(acyl groups) including (1-6C)alkanoyl groups, such as formyl, acetyland the like; arylmethyl groups, such as benzyl (Bn), p-methoxybenzyl(PMB), 9-fluorenylmethyl (Fm), diphenylmethyl (benzhydryl, DPM) and thelike. Additionally, two hydroxyl groups can also be protected as analkylidene group, such as prop-2-ylidine, formed, for example, byreaction with a ketone, such as acetone.

General Synthetic Procedures

The compounds of this invention can be prepared from readily availablestarting materials using the following general methods and procedures orby using other information readily available to those of ordinary skillin the art. Although a particular embodiment of the present inventionmay be shown or described herein, those skilled in the art willrecognize that all embodiments or aspects of the present invention canbe prepared using the methods described herein or by using othermethods, reagents and starting materials known to those skilled in theart. It will also be appreciated that where typical or preferred processconditions (i.e., reaction temperatures, times, mole ratios ofreactants, solvents, pressures, etc.) are given, other processconditions can also be used unless otherwise stated. While the optimumreaction conditions may vary depending on the particular reactants orsolvent used, such conditions can be readily determined by one skilledin the art by routine optimization procedures.

Additionally, as will be apparent to those skilled in the art,conventional protecting groups may be necessary or desired to preventcertain functional groups from undergoing undesired reactions. Thechoice of a suitable protecting group for a particular functional groupas well as suitable conditions for protection and deprotection of suchfunctional groups are well-known in the art. Protecting groups otherthan those illustrated in the procedures described herein may be used,if desired. For example, numerous protecting groups, and theirintroduction and removal, are described in T. W. Greene and G. M. Wuts,Protecting Groups in Organic Synthesis, Third Edition, Wiley, New York,1999, and references cited therein.

By way of illustration, the compounds of this invention can be preparedby a process comprising:

(a) reacting a compound of formula 1:

or a salt thereof, with a compound of formula 2:

wherein X¹ represents a leaving group, and P¹ and p² each independentlyrepresent a hydrogen atom or a hydroxyl-protecting group;

(b) reacting a compound of formula 3:

or salt thereof; with a compound of formula 4:

wherein X² represents a leaving group, and P³ and p⁴ each independentlyrepresent a hydrogen atom or a hydroxyl-protecting group;

(c) coupling a compound of formula 5:

with a compound of formula 6:

wherein X^(Qa) and X^(Qb) each independently represent functional groupsthat couple to form a group Q, P^(5a) represents a hydrogen atom or anamino-protecting group; and P^(5b) and P⁶ each independently represent ahydrogen atom or a hydroxyl-protecting group;

(d) for a compound of formula I wherein R⁵ represents a hydrogen atom,reacting a compound of formula 3 with a compound of formula 7:

or a hydrate thereof (e.g., a glyoxal), in the presence of a reducingagent, wherein P⁷ represents a hydrogen atom or a hydroxyl-protectinggroup;

(e) reacting a compound of formula I with a compound of formula 8:

or a hydrate thereof, in the presence of a reducing agent, wherein P⁸and P⁹ each independently represent a hydrogen atom or ahydroxyl-protecting group, P¹⁰ represents a hydrogen atom or anamino-protecting group, and R⁴ represents a residue that, together withthe carbon to which it is attached, affords a group R⁴ upon completionof the reaction;

(f) reacting a compound of formula 9:

wherein X³ represents a leaving group, with a compound of formula 10:

wherein P¹¹ and P¹² each independently represent a hydrogen atom or ahydroxyl-protecting group, and P¹³ represents a hydrogen atom or anamino-protecting group; or

(g) reacting a compound of formula 11:

or a hydrate thereof; wherein R⁴ represents a residue that, togetherwith the carbon to which it is attached, affords a group R⁴ uponcompletion of the reaction; with a compound of formula 10 in thepresence of a reducing agent;

(h) reacting a compound of formula 12:

wherein Y¹ represents chloro, bromo, iodo or CF₃SO₂O—, P¹⁴ represents ahydrogen atom or an amino-protecting group; and P¹⁵ and P¹⁶ eachindependently represent a hydrogen atom or a hydroxyl-protecting group;with a compound of the formula: R¹—B(OH)₂ in the presence of a couplingcatalyst; and then

removing any protecting group P¹, P^(2, P) ³, P⁴, P^(5a), P^(5b), P⁶,P⁷, P⁸, P⁹, P¹⁰, P¹¹, P¹², P¹³, P¹⁴, P¹⁵ and P¹⁶ to provide a compoundof formula 1; and optionally, forming a pharmaceutically acceptable saltthereof.

Generally, if a salt of one of the starting materials is used in theprocesses described above, such as an acid addition salt, the salt istypically neutralized before or during the reaction process. Thisneutralization reaction is typically accomplished by contacting the saltwith one molar equivalent of a base for each molar equivalent of acidaddition salt.

In process (a), i.e., the reaction between the compounds of formula 1and 2, the leaving group represented by X¹ can be, for example, halo,such as chloro, bromo or iodo, or a sulfonic ester group, such asmesylate or tosylate. The groups P¹ and P² can be, for example,trimethylsilyl and benzyl, respectively. This reaction is typicallyconducted in an inert diluent, such as acetonitrile, in the presence ofa base. For example, this reaction can be conducted in the presence of atertiary amine, such as diisopropylethylamine. Generally, this reactionis conducted at a temperature in the range of from 0° C. to 100° C.until the reaction is substantially complete. The reaction product isthen isolated using conventional procedures, such as extraction,recrystallization, chromatography and the like.

Compounds of formula 1 are generally known in the art or can be preparedfrom commercially available starting materials and reagents usingwell-known procedures. For example, compounds of formula 1 can beprepared by reacting an isocyanate compound of formula 13:

with a compound of formula 14:

wherein P¹⁷ represents an amino-protecting group, such as a benzylgroup; followed by removal of the protecting group P¹⁷ to afford acompound of formula 1. For example, when P¹⁷ is a benzyl group, thebenzyl group is removed by hydrogenolysis, for example, using hydrogenand a group VIII metal catalyst, such as palladium on carbon. When Wrepresents NW^(a), the hydrogenolysis reaction is conveniently performedusing Pearlman's catalyst (i.e., Pd(OH)₂).

Compounds of formula 2 can be prepared by various procedures describedherein or by procedures that are well-known to those skilled in the art.For example, the hydroxyl group of a compound of formula 22 below, canbe readily converted into a leaving group using well-known reagents andprocedures. By way of illustration, a hydroxyl group can be convertedinto a halo group using an inorganic acid halide, such as thionylchloride, phosphorous trichloride, phosphorous tribromide, phosphorousoxychloride and the like, or a halogen acid, such a hydrogen bromide.

In process (b), i.e., the reaction of a compound of formula 3 with acompound of formula 4, the leaving represented by X² can be, forexample, halo, such as chloro, bromo or iodo, or a sulfonic ester group,such as mesylate or tosylate. The groups P³ and P⁴ can be, for example,tert-butyldimethylsilyl and benzyl, respectively. This reaction istypically conducted in the presence of a base, such as sodiumbicarbonate, and an alkali metal iodide, such as sodium iodide.Generally, this reaction is conducted in an inert diluent, such astetrahydrofuran, at a temperature ranging from 25° C. to 100° C. untilthe reaction is substantially complete. The reaction product is thenisolated using conventional procedures, such as extraction,recrystallization, chromatography and the like.

Compounds of formula 3 can be prepared by reacting a compound of formula1 with a compound of formula 15:X⁴—R⁴—NP¹⁸P¹⁹  15

wherein X⁴ represents a leaving group such as halo, such as chloro,bromo or iodo, or sulfonic ester group, such as mesylate or tosylate.This reaction is typically conducted by contacting a compound of formula1 with a compound of formula 15 in an inert diluent, such asacetonitrile, DMF or mixtures thereof, at a temperature ranging fromabout 0° C. to about 100° C. until the reaction is substantiallycomplete. Subsequent removal of the protecting groups, P¹⁸ and P¹⁹,affords a compound of formula 3. For example, when P¹⁸ is atert-butoxycarbonyl group and P¹⁹ is hydrogen, the tert-butoxycarbonylgroup can be removed by treating the protected compound withtrifluoroacetic acid.

Alternatively, compounds of formula 3 can be obtained by reductiveamination of a compound of formula 11. The reductive amination can beperformed by reacting the compound of formula 11 with, for example,benzylamine and hydrogen in the presence of palladium on carbon.

Compounds of formula 11 may be prepared by oxidizing the correspondingalcohol of formula 16:

using a suitable oxidizing agent, such as sulfur trioxide pyridinecomplex and dimethyl sulfoxide. This oxidation reaction is typicallyconducted in an inert diluent, such as dichloromethane, the presence ofa tertiary amine, such as diisopropylethylamine, at a temperatureranging from about −20° C. to about 25° C.

Compounds of formula 16 can be prepared by reacting a compound offormula 1 with a compound of formula 17:X⁵—R⁴—OH  17wherein X⁵ represents a leaving group such as halo, such as chloro,bromo or iodo, or a sulfonic ester group, such as mesylate or tosylate.

Compounds of formula 4 can be prepared by reacting a compound of formula18:

with a reducing agent, such as borane. If desired, such a reduction canbe performed in the presence of a chiral catalyst to provide compoundsof formula 4 in chiral form. For example, compounds of formula 18 can bereduced in the presence of a chiral catalyst formed from(R)-(+)-α,α-diphenyl-2-pyrrolidinemethanol and trimethylboroxine; oralternatively, from (S)-(−)-(α,α-diphenyl-2-pyrrolidinemethanol andtrimethylboroxine. The resulting hydroxyl group can then be protectedwith a hydroxyl-protecting group, P³, by reaction with, for example,tert-butyldimethylsilyl trifluoromethanesulfonate.

Compounds of formula 18 in which X² represents a bromine atom can beprepared by reacting a compound of formula 19:

with bromine in the presence of a Lewis acid, such as boron trifluoridediethyl etherate. Compounds of formula 19 are well-known in the art orcan be prepared by well-known procedures using commercially availablestarting materials and reagents.

Referring to process (c), i.e., the reaction of a compound of formula 5with a compound of formula 6, it will be appreciated that the groupsX^(Qa) and X^(Qb) should be selected so as to afford the desired group Qupon completion of the reaction. For example, when the desired group Qis an amide group, i.e., —N(Q^(a))C(O)— or —C(O)N(Q^(b)), one of X^(Qa)and X^(Qb) can be an amine group (i.e., —NHQ^(a) or —NHQ^(b)) and theother can be a carboxyl group (i.e., —COOH) or a reactive derivativethereof (such as acyl halide, such as an acyl chloride or acyl bromide).The groups P^(5a), P^(5b) and P⁶ can be, for example, benzyl,trimethylsilyl and benzyl, respectively. When Q is an amide group, thereaction can be performed under conventional amide coupling conditions.Similarly, when the desired group Q is a sulfonamide, i.e.,—N(Q^(c))S(O)₂— or —S(O)₂N(Q^(d))—, one of X^(Qa) and X^(Qb) can be anamine group, —NHQ^(c) or —NHQ^(d) and the other can be a sulfonyl halidegroup (such as sulfonyl chloride or sulfonyl bromide).

Compounds of formula 5 can be prepared by reacting a compound of formula1 with a compound of formula 20:X⁶—(R^(4a))_(d)-(A¹)_(e)-(R^(4b))_(f)—X^(Qa′)  20wherein X⁶ represents a leaving group including halo, such as chloro,bromo or iodo, and a sulfonic ester group, such as mesylate or tosylate;and X^(Qa′) represents X^(Qa), such as a carboxyl group or an aminogroup NHQ^(a), or a protected derivative thereof, such as a(1-6C)alkoxycarbonylamino group or a tert-butoxycarbonylamino group.This reaction is typically conducted by a method analogous to that usedto prepare compounds of formula 3, followed by removing any protectinggroup in X^(Qa′).

Compounds of formula 6 can be prepared by reacting a compound of formula10 with a compound of formula 21:X^(Qb′)—(R^(4c))_(g)-(A²)_(h)-(R^(4d))_(i)—X⁷  21wherein X⁷ represents a leaving group including halo, such as chloro,bromo or iodo, and a sulfonic ester group, such as mesylate or tosylate;and X^(Qb′) represents X^(Qb), such as a carboxyl group or an aminogroup NHQ^(b), or a protected derivative thereof, such as a(1-6C)alkoxycarbonyl group or a tert-butoxycarbonylamino group. Thisreaction is typically conducted by a method analogous to that used toprepare compounds of formula 3, followed by removing any protectinggroup in X^(Qb′).

Referring to process (d), i.e., the reaction of a compound of formula 3with a compound of formula 7, any suitable reducing agent may be used inthis reaction. For example, the reducing agent can be hydrogen in thepresence of a Group VIII metal catalyst, such as palladium on carbon; ora metal hydride reagent, such as sodium triacetoxyborohydride. The groupP⁷ can be, for example, benzyl. This reaction is typically conducted inan inert diluent and a protic solvent, such as a mixture ofdichloroethane and methanol, at a temperature in the range of from 0° C.to 100° C. until the reaction is substantially complete.

Compounds of formula 7 in the form of a hydrate can be prepared byconventional procedures, for example, by dibrominating a compound offormula 19, and then hydrolyzing the resulting dibromide to form aglyoxal or a hydrate thereof. For example, a compound of formula 19 canbe reacted with hydrogen bromide and then hydrolyzed with water to formthe corresponding glyoxal hydrate.

Referring to process (e), i.e., the reaction of a compound of formula 1with a compound of formula 8, any suitable reducing agent may be used inthis reaction. For example, the reducing agent may be hydrogen in thepresence of a Group VIII metal catalyst, such as palladium on carbon; ora metal hydride reagent, such as sodium triacetoxyborohydride. Thegroups P⁸, P⁹ and P¹⁰ can be, for example, trimethylsilyl, benzyl andbenzyl, respectively. Typically, this reduction reaction is conducted inan inert diluent and a protic solvent, such as dichloroethane andmethanol, at a temperature in the range of from 0° C. to 100° C. untilthe reaction is substantially complete.

Compounds of formula 8 may be prepared by oxidizing a compound offormula 22:

using any suitable oxidizing agent, such as sulfur trioxide pyridinecomplex and dimethyl sulfoxide. This reaction is typically conducted inthe presence of a tertiary amine, such as diisopropylethylamine, at atemperature in the range of from about −20° C. to about 25° C. until theoxidation is substantially complete.

Compounds of formula 22 can be prepared by reacting a compound offormula 10 with a compound of formula 23:HO—R⁴—X⁸  23wherein X⁸ represents a leaving group including halo, such as chloro,bromo or iodo, and a sulfonic ester group, such as mesylate or tosylate.

Referring to process (f), i.e., the reaction of a compound of formula 9with a compound of formula 10, the leaving group represented by X³ canbe, for example, halo, such as chloro, bromo or iodo, or a sulfonicester group, such as mesylate or tosylate. The groups P¹¹, P¹² and P¹³can be, for example, trimethylsilyl, benzyl and benzyl, respectively.This reaction is typically conducted an inert diluent, such asacetonitrile, in the presence of a suitable base. For example, thisreaction can be conducted in the presence of a tertiary amine, such asdiisopropylethylamine. Generally, this reaction is conducted at atemperature in the range of from 0° C. to 100° C. until the reaction issubstantially complete.

Compounds of formula 9 can be prepared by steps analogous to those ofmethods (a) to (e) herein, starting from a compound of formula 1.Additionally, compounds of formula 10 can be prepared from compounds offormula 4 by reaction with an amine of formula P¹³NH₂.

Referring to process (g), i.e., the reaction of a compound of formula 11with a compound of formula 10, any suitable reducing agent may be usedin this reaction. For example, the reducing agent may be hydrogen in thepresence of a Group VIII metal catalyst, such as palladium on carbon; ora metal hydride reagent, such as sodium triacetoxyborohydride. Thegroups P¹¹, P¹² and P¹³ can be, for example, tert-butyldimethylsilyl,benzyl and benzyl, respectively. Typically, this reduction reaction isconducted in an inert diluent and a protic solvent, such asdichloroethane and methanol, at a temperature in the range of from 0° C.to 100° C. until the reaction is substantially complete.

Compounds of formula 11 are readily prepared by oxidation of thecorresponding alcohol or by hydrolysis of the corresponding acetal. Anysuitable oxidizing agent may be employed in this reaction to provide thealdehyde, such as sulfur trioxide pyridine complex and dimethylsulfoxide. The acetal may be hydrolyzed under conventional conditionsusing aqueous acid to provide the aldehyde.

Referring to process (h), i.e., the reaction of a compound of formula 12with a boronic acid compound of formula R¹—B(OH)₂, any suitable couplingcatalyst can be used to couple 12 with the boronic acid derivative usingSuzuki coupling reaction conditions. For example, representativecatalysts include palladium and nickel catalysts, such as[1,1′-bis(diphenylphosphino)ferrocene]dicloropalladium(II),bis[1,2-bis(diphenylphosphino)propane]palladium(O), palladium(II)acetate, [1,1′-bis(diphenylphosphino)ferrocene]dicloronickel(II) and thelike. Optionally, a base is employed in this reaction, such as sodiumcarbonate, sodium bicarbonate, potassium phosphate, triethylamine andthe like. Generally, compounds of formula 12 in which Y¹ representsbromo, iodo or CF₃SO₂O— (i.e., triflate) are more reactive in thecoupling reaction and therefore, such compounds are typically preferred.Particularly useful compounds of formula 12 are those in which Y¹ isbromo. In the compounds of formula 12, P¹⁴, P¹⁵ and P¹⁶ can be anysuitable amino- and hydroxyl-protecting groups, such as benzyl,tert-butyldimethylsilyl and benzyl, respectively.

The boronic acid compounds employed in process (h) are eithercommercially available or can be prepared from commercially availablestarting materials and reagents using procedures well known to thoseskilled in the art. For example, boronic acid derivatives can beprepared by reacting the corresponding lithium derivative with atrialkyl borate, such as triisopropyl borate, and then hydrolyzing theresulting product to provide the boronic acid derivative.

Those skilled in the art will appreciate that the Suzuki couplingreaction can be conducted at any suitable step of the synthetic process.For example, the Suzuki coupling reaction can be conducted as the finalstep of the synthetic process after the remaining portions of thecompound have been prepared. Alternatively, the Suzuki coupling reactioncan be conducted earlier in the synthetic process to prepareintermediates having the desired biaryl ring system. Additionally, theboronic acid moiety may be present on the compound of formula 12, i.e.,where Y¹ in 12 is —B(OH)₂; and this compound may be coupled to acompound of the formula: R¹—Y², where Y² represents chloro, bromo, iodoor CF₃SO₂O—.

Compounds of formula I in which R⁶ and R⁷ together form—NR^(7g)C(O)—CR^(7h)R^(7i)—CR^(7j)R^(7k)— or—CR^(7l)R^(7m)—CR^(7n)R^(7o)—C(O) —NR^(7p)— may be prepared by reducinga corresponding compound of formula I in which R⁶ and R⁷ together form—NR^(7a)C(O)—CR^(7b)═CR^(7c)— or —CR^(7d)═CR^(7e)—C(O)—NR^(7f)—. Forexample, such compounds can be reduced by catalytic hydrogenation in thepresence of a suitable hydrogenation catalyst.

Further details regarding specific reaction conditions and otherprocedures for preparing representative compounds of this invention orintermediates thereof are described in the Examples set forth below.

Pharmaceutical Compositions and Formulations

The compounds of this invention are typically administered to a patientin the form of a pharmaceutical composition or formulation. Suchpharmaceutical compositions may be administered to the patient by anyacceptable route of administration including, but not limited to,inhaled, oral, nasal, topical (including transdermal) and parenteralmodes of administration. It will be understood that any form of thecompounds of this invention, (i.e., free base, pharmaceuticallyacceptable salt, solvate, etc.) that is suitable for the particular modeof administration can be used in the pharmaceutical compositionsdiscussed herein.

Accordingly, in one of its compositions aspects, this invention isdirected to a pharmaceutical composition comprising a pharmaceuticallyacceptable carrier or excipient and a therapeutically effective amountof a compound of formula I, or a pharmaceutically acceptable saltthereof. Optionally, such pharmaceutical compositions may contain othertherapeutic and/or formulating agents if desired.

The pharmaceutical compositions of this invention typically contain atherapeutically effective amount of a compound of the present inventionor a pharmaceutically acceptable salt thereof. Typically, suchpharmaceutical compositions will contain from about 0.01 to about 95% byweight of the active agent; including, from about 0.01 to about 30% byweight; such as from about 0.01 to about 10% by weight of the activeagent.

Any conventional carrier or excipient may be used in the pharmaceuticalcompositions of this invention. The choice of a particular carrier orexcipient, or combinations of carriers or exipients, will depend on themode of administration being used to treat a particular patient or typeof medical condition or disease state. In this regard, the preparationof a suitable pharmaceutical composition for a particular mode ofadministration is well within the scope of those skilled in thepharmaceutical arts. Additionally, the ingredients for such compositionsare commercially available from, for example, Sigma, P.O. Box 14508, St.Louis, Mo. 63178. By way of further illustration, conventionalformulation techniques are described in Remington: The Science andPractice of Pharmacy, 20^(th) Edition, Lippincott Williams & White,Baltimore, Md. (2000); and

H. C. Ansel et al., Pharmaceutical Dosage Forms and Drug DeliverySystems, 7^(th) Edition, Lippincott Williams & White, Baltimore, Md.(1999).

Representative examples of materials which can serve as pharmaceuticallyacceptable carriers include, but are not limited to, the following: (1)sugars, such as lactose, glucose and sucrose; (2) starches, such as cornstarch and potato starch; (3) cellulose, and its derivatives, such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;(4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8)excipients, such as cocoa butter and suppository waxes; (9) oils, suchas peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil,corn oil and soybean oil; (10) glycols, such as propylene glycol; (11)polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol;(12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14)buffering agents, such as magnesium hydroxide and aluminum hydroxide;(15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18)Ringer's solution; (I 9) ethyl alcohol; (20) phosphate buffer solutions;(21) compressed propellant gases, such as chlorofluorocarbons andhydrofluorocarbons; and (22) other non-toxic compatible substancesemployed in pharmaceutical compositions.

The pharmaceutical compositions of this invention are typically preparedby thoroughly and intimately mixing or blending a compound of theinvention with a pharmaceutically acceptable carrier and one or moreoptional ingredients. If necessary or desired, the resulting uniformlyblended mixture can then be shaped or loaded into tablets, capsules,pills, canisters, cartridges, dispensers and the like using conventionalprocedures and equipment.

In one embodiment, the pharmaceutical compositions of this invention aresuitable for inhaled administration. Suitable pharmaceuticalcompositions for inhaled administration will typically be in the form ofan aerosol or a powder. Such compositions are generally administeredusing well-known delivery devices, such as a nebulizer inhaler, ametered-dose inhaler (MDI), a dry powder inhaler (DPI) or a similardelivery device.

In a specific embodiment of this invention, the pharmaceuticalcomposition comprising the active agent is administered by inhalationusing a nebulizer inhaler. Such nebulizer devices typically produce astream of high velocity air that causes the pharmaceutical compositioncomprising the active agent to spray as a mist that is carried into thepatient's respiratory tract. Accordingly, when formulated for use in anebulizer inhaler, the active agent is typically dissolved in a suitablecarrier to form a solution. Alternatively, the active agent can bemicronized and combined with a suitable carrier to form a suspension ofmicronized particles of respirable size, where micronized is typicallydefined as having about 90% or more of the particles with a diameter ofless than about 10 μm. Suitable nebulizer devices are providedcommercially, for example, by PARI GmbH (Starnberg, German). Othernebulizer devices include Respimat (Boehringer Ingelheim) and thosedisclosed, for example, in U.S. Pat. No. 6,123,068 and WO 97/12687.

A representative pharmaceutical composition for use in a nebulizerinhaler comprises an isotonic aqueous solution comprising from about0.05 μg/mL to about 10 mg/mL of a compound of formula I or apharmaceutically acceptable salt or solvate or stereoisomer thereof.

In another specific embodiment of this invention, the pharmaceuticalcomposition comprising the active agent is administered by inhalationusing a dry powder inhaler. Such dry powder inhalers typicallyadminister the active agent as a free-flowing powder that is dispersedin a patient's air-stream during inspiration. In order to achieve afree-flowing powder, the active agent is typically formulated with asuitable excipient such as lactose or starch.

A representative pharmaceutical composition for use in a dry powderinhaler comprises dry lactose having a particle size between about 1 μmand about 100 μm and micronized particles of a compound of formula I, ora pharmaceutically acceptable salt or solvate or stereoisomer thereof.

Such a dry powder formulation can be made, for example, by combining thelactose with the active agent and then dry blending the components.Alternatively, if desired, the active agent can be formulated without anexcipient. The pharmaceutical composition is then typically loaded intoa dry powder dispenser, or into inhalation cartridges or capsules foruse with a dry powder delivery device.

Examples of dry powder inhaler delivery devices include Diskhaler(GlaxoSmithKline, Research Triangle Park, N.C.) (see, e.g., U.S. Pat.No. 5,035,237); Diskus (GlaxoSmithKline) (see, e.g., U.S. Pat. No.6,378,519; Turbuhaler (AstraZeneca, Wilmington, Del.) (see, e.g., U.S.Pat. No. 4,524,769); Rotahaler (GlaxoSmithKline) (see, e.g., U.S. Pat.No. 4,353,365) and Handihaler (Boehringer Ingelheim). Further examplesof suitable DPI devices are described in U.S. Pat. Nos. 5,415,162,5,239,993, and 5,715,810 and references cited therein.

In yet another specific embodiment of this invention, the pharmaceuticalcomposition comprising the active agent is administered by inhalationusing a metered-dose inhaler. Such metered-dose inhalers typicallydischarge a measured amount of the active agent or a pharmaceuticallyacceptable salt thereof using compressed propellant gas. Accordingly,pharmaceutical compositions administered using a metered-dose inhalertypically comprise a solution or suspension of the active agent in aliquefied propellant. Any suitable liquefied propellant may be employedincluding chlorofluorocarbons, such as CCl₃F, and hydrofluoroalkanes(HFAs), such as 1,1,1,2-tetrafluoroethane (HFA 134a) and1,1,1,2,3,3,3-heptafluoro-n-propane, (HFA 227). Due to concerns aboutchlorofluorocarbons affecting the ozone layer, formulations containingHFAs are generally preferred. Additional optional components of HFAformulations include co-solvents, such as ethanol or pentane, andsurfactants, such as sorbitan trioleate, oleic acid, lecithin, andglycerin. See, for example, U.S. Pat. No. 5,225,183, EP 0717987 A2, andWO 92/22286.

A representative pharmaceutical composition for use in a metered-doseinhaler comprises from about 0.01% to about 5% by weight of a compoundof formula I, or a pharmaceutically acceptable salt or solvate orstereoisomer thereof; from about 0% to about 20% by weight ethanol; andfrom about 0% to about 5% by weight surfactant; with the remainder beingan HFA propellant.

Such compositions are typically prepared by adding chilled orpressurized hydrofluoroalkane to a suitable container containing theactive agent, ethanol (if present) and the surfactant (if present). Toprepare a suspension, the active agent is micronized and then combinedwith the propellant. The formulation is then loaded into an aerosolcanister, which forms a portion of a metered-dose inhaler device.Examples of metered-dose inhaler devices developed specifically for usewith HFA propellants are provided in U.S. Pat. Nos. 6,006,745 and6,143,277. Alternatively, a suspension formulation can be prepared byspray drying a coating of surfactant on micronized particles of theactive agent. See, for example, WO 99/53901 and WO 00/61108.

For additional examples of processes of preparing respirable particles,and formulations and devices suitable for inhalation dosing see U.S.Pat. Nos. 6,268,533, 5,983,956, 5,874,063, and 6,221,398, and WO99/55319 and WO 00/30614.

In another embodiment, the pharmaceutical compositions of this inventionare suitable for oral administration. Suitable pharmaceuticalcompositions for oral administration may be in the form of capsules,tablets, pills, lozenges, cachets, dragees, powders, granules; or as asolution or a suspension in an aqueous or non-aqueous liquid; or as anoil-in-water or water-in-oil liquid emulsion; or as an elixir or syrup;and the like; each containing a predetermined amount of a compound ofthe present invention as an active ingredient.

When intended for oral administration in a solid dosage form (i.e., ascapsules, tablets, pills and the like), the pharmaceutical compositionsof this invention will typically comprise a compound of the presentinvention as the active ingredient and one or more pharmaceuticallyacceptable carriers, such as sodium citrate or dicalcium phosphate.Optionally or alternatively, such solid dosage forms may also comprise:(1) fillers or extenders, such as starches, lactose, sucrose, glucose,mannitol, and/or silicic acid; (2) binders, such ascarboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone,sucrose and/or acacia; (3) humectants, such as glycerol; (4)disintegrating agents, such as agar-agar, calcium carbonate, potato ortapioca starch, alginic acid, certain silicates, and/or sodiumcarbonate; (5) solution retarding agents, such as paraffin; (6)absorption accelerators, such as quaternary ammonium compounds; (7)wetting agents, such as cetyl alcohol and/or glycerol monostearate; (8)absorbents, such as kaolin and/or bentonite clay; (9) lubricants, suchas talc, calcium stearate, magnesium stearate, solid polyethyleneglycols, sodium lauryl sulfate, and/or mixtures thereof; (10) coloringagents; and (11) buffering agents.

Release agents, wetting agents, coating agents, sweetening, flavoringand perfuming agents, preservatives and antioxidants can also be presentin the pharmaceutical compositions of this invention. Examples ofpharmaceutically acceptable antioxidants include: (1) water-solubleantioxidants, such as ascorbic acid, cysteine hydrochloride, sodiumbisulfate, sodium metabisulfate sodium sulfite and the like; (2)oil-soluble antioxidants, such as ascorbyl palmitate, butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propylgallate, alpha-tocopherol, and the like; and (3) metal-chelating agents,such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol,tartaric acid, phosphoric acid, and the like. Coating agents fortablets, capsules, pills and like, include those used for entericcoatings, such as cellulose acetate phthalate (CAP), polyvinyl acetatephthalate (PVAP), hydroxypropyl methylcellulose phthalate, methacrylicacid-methacrylic acid ester copolymers, cellulose acetate trimellitate(CAT), carboxymethyl ethyl cellulose (CMEC), hydroxypropyl methylcellulose acetate succinate (HPMCAS), and the like.

If desired, the pharmaceutical compositions of the present invention mayalso be formulated to provide slow or controlled release of the activeingredient using, by way of example, hydroxypropyl methyl cellulose invarying proportions; or other polymer matrices, liposomes and/ormicrospheres.

In addition, the pharmaceutical compositions of the present inventionmay optionally contain opacifying agents and may be formulated so thatthey release the active ingredient only, or preferentially, in a certainportion of the gastrointestinal tract, optionally, in a delayed manner.Examples of embedding compositions which can be used include polymericsubstances and waxes. The active ingredient can also be inmicro-encapsulated form, if appropriate, with one or more of theabove-described excipients.

Suitable liquid dosage forms for oral administration include, by way ofillustration, pharmaceutically acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. Such liquid dosage formstypically comprise the active ingredient and an inert diluent, such as,for example, water or other solvents, solubilizing agents andemulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate,ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol,1,3-butylene glycol, oils (esp., cottonseed, groundnut, corn, germ,olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol,polyethylene glycols and fatty acid esters of sorbitan, and mixturesthereof. Suspensions, in addition to the active ingredient, may containsuspending agents such as, for example, ethoxylated isostearyl alcohols,polyoxyethylene sorbitol and sorbitan esters, microcrystallinecellulose, aluminium metahydroxide, bentonite, agar-agar and tragacanth,and mixtures thereof.

When intended for oral administration, the pharmaceutical compositionsof this invention are preferably packaged in a unit dosage form. Theterm “unit dosage form” means a physically discrete unit suitable fordosing a patient, i.e., each unit containing a predetermined quantity ofactive agent calculated to produce the desired therapeutic effect eitheralone or in combination with one or more additional units. For example,such unit dosage forms may be capsules, tablets, pills, and the like.

The compounds of this invention can also be administered transdermallyusing known transdermal delivery systems and excipents. For example, acompound of this invention can be admixed with permeation enhancers,such as propylene glycol, polyethylene glycol monolaurate,azacycloalkan-2-ones and the like, and incorporated into a patch orsimilar delivery system. Additional excipients including gelling agents,emulsifiers and buffers, may be used in such transdermal compositions ifdesired.

The pharmaceutical compositions of this invention may also contain othertherapeutic agents that are co-administered with a compound of formulaI, or pharmaceutically acceptable salt or solvate or stereoisomerthereof. For example, the pharmaceutical compositions of this inventionmay further comprise one or more therapeutic agents selected from otherbronchodilators (e.g., PDE₃ inhibitors, adenosine 2b modulators and β₂adrenergic receptor agonists); anti-inflammatory agents (e.g. steroidalanti-inflammatory agents, such as corticosteroids; non-steroidalanti-inflammatory agents (NSAIDs), and PDE₄ inhibitors); othermuscarinic receptor antagonists (i.e., antichlolinergic agents);antiinfective agents (e.g. Gram positive and Gram negative antibioticsor antivirals); antihistamines; protease inhibitors; and afferentblockers (e.g., D₂ agonists and neurokinin modulators). The othertherapeutic agents can be used in the form of pharmaceuticallyacceptable salts or solvates. Additionally, if appropriate, the othertherapeutic agents can be used as optically pure stereoisomers.

Representative β₂ adrenergic receptor agonists that can be used incombination with, and in addition to, the compounds of this inventioninclude, but are not limited to, salmeterol, salbutamol, formoterol,salmefamol, fenoterol, terbutaline, albuterol, isoetharine,metaproterenol, bitolterol, pirbuterol, levalbuterol and the like, orpharmaceutically acceptable salts thereof. Other β₂ adrenergic receptoragonists that can be used in combination with the compounds of thisinvention include, but are not limited to,3-(4-{[6-({(2R)-2-hydroxy-2-[4-hydroxy-3-(hydroxymethyl)phenyl]ethyl}amino)-hexyl]oxy}butyl)benzenesulfonamideand3-(-3-{[7-({(2R)-2-hydroxy-2-[4-hydroxy-3-(hydroxymethyl)phenyl]ethyl}amino)heptyl]oxy}propyl)benzenesulfonamideand related compounds disclosed in WO 02/066422, published on Aug. 29,2002;3-[3-(4-{[6-([(2R)-2-hydroxy-2-[4-hydroxy-3-(hydroxymethyl)phenyl]ethyl}amino)hexyl]oxy}butyl)-phenyl]imidazolidine-2,4-dioneand related compounds disclosed in WO 02/070490, published Sep. 12,2002;3-(4-{[6-({(2R)-2-[3-(formylamino)-4-hydroxyphenyl]-2-hydroxyethyl}amino)hexyl]oxy}butyl)-benzenesulfonamide,3-(4-{[6-({(2S)-2-[3-(formylamino)-4-hydroxyphenyl]-2-hydroxyethyl}amino)hexyl]oxy}butyl)-benzenesulfonamide,3-(4-{[6-({(2R/S)-2-[3-(formylamino)-4-hydroxyphenyl]-2-hydroxyethyl}amino)hexyl]oxy}butyl)-benzenesulfonamide,N-(tert-butyl)-3-(4-{[6-({(2R)-2-[3-(formylamino)-4-hydroxyphenyl]-2-hydroxyethyl}amino)hexyl]-oxy}butyl)benzenesulfonamide,N-(tert-butyl)-3-(4-{[6-({(2S)-2-[3-(formylamino)-4-hydroxyphenyl]-2-hydroxyethyl}amino)-hexyl]oxy}butyl)-benzenesulfonamide,N-(tert-butyl)-3-(4-{[6-({(2R/S)-2-[3-(formylamino)-4-hydroxyphenyl]-2-hydroxyethyl}amino)hexyl]-oxy}butyl)benzenesulfonamideand related compounds disclosed in WO 02/076933, published on Oct. 3,2002;4-{(1R)-2-[(6-{2-[(2,6-dichlorobenzyl)oxy]ethoxy}hexyl)amino]-1-hydroxyethyl}-2-(hydroxymethyl)phenoland related compounds disclosed in WO 03/024439, published on Mar. 27,2003; and pharmaceutically acceptable salts thereof. When employed, theβ₂-adrenoreceptor agonist will be present in the pharmaceuticalcomposition in a therapeutically effective amount. Typically, theβ₂-adrenoreceptor agonist will be present in an amount sufficient toprovide from about 0.05 μg to about 500 μg per dose.

Representative steroidal anti-inflammatory agents that can be used incombination with the compounds of this invention include, but are notlimited to, methyl prednisolone, prednisolone, dexamethasone,fluticasone propionate,6,9-difluoro-17-[(2-furanylcarbonyl)oxy]-11-hydroxy-16-methyl-3-oxoandrosta-1,4-diene-17-carbothioicacid S-fluoromethyl ester,6,9-difluoro-11-hydroxy-16-methyl-3-oxo-17-propionyloxy-androsta-1,4-diene-17-carbothioicacid S-(2-oxo-tetrahydrofuran-3S-yl) ester, beclomethasone esters (e.g.the 17-propionate ester or the 17,21-dipropionate ester), budesonide,flunisolide, mometasone esters (e.g. the furoate ester), triamcinoloneacetonide, rofleponide, ciclesonide, butixocort propionate, RPR-106541,ST-126 and the like, or pharmaceutically-acceptable salts thereof. In aparticular embodiment, the steroidal anti-inflammatory agent is6α,9α-difluoro-17α-[(2-furanylcarbonyl)oxy]-11β-hydroxy-16α-methyl-3-oxoandrosta-1,4-diene-17β-carbothioicacid S-fluoromethyl ester or a pharmaceutically acceptable salt orsolvate thereof. When employed, the steroidal anti-inflammatory agentwill be present in the pharmaceutical composition in a therapeuticallyeffective amount. Typically, the steroidal anti-inflammatory agent willbe present in an amount sufficient to provide from about 0.05 μg toabout 500 μg per dose.

Other suitable combinations include, for example, otheranti-inflammatory agents, e.g., NSAIDs (such as sodium cromoglycate;nedocromil sodium; phosphodiesterase (PDE) inhibitors (e.g.theophylline, PDE4 inhibitors or mixed PDE3/PDE4 inhibitors);leukotriene antagonists (e.g. monteleukast); inhibitors of leukotrienesynthesis; iNOS inhibitors; protease inhibitors, such as tryptase andelastase inhibitors; beta-2 integrin antagonists and adenosine receptoragonists or antagonists (e.g. adenosine 2a agonists); cytokineantagonists (e.g. chemokine antagonists such as, an interleukin antibody(IL antibody), specifically, an IL-4 therapy, an IL-13 therapy, or acombination thereof); or inhibitors of cytokine synthesis.

For example, representative phosphodiesterase-4 (PDE4) inhibitors ormixed PDE3/PDE4 inhibitors that can be used in combination with thecompounds of this invention include, but are not limited to cis4-cyano-4-(3-cyclopentyloxy-4-methoxyphenyl)cyclohexan-1-carboxylicacid,2-carbomethoxy-4-cyano-4-(3-cyclopropylmethoxy-4-difluoromethoxyphenyl)cyclohexan-1-one;cis-[4-cyano-4-(3-cyclopropylmethoxy-4-difluoromethoxyphenyl)cyclohexan-1-ol];cis-4-cyano-4-[3-(cyclopentyloxy)-4-methoxyphenyl]cyclohexane-1-carboxylicacid and the like, or pharmaceutically acceptable salts thereof. Otherrepresentative PDE4 or mixed PDE4/PDE3 inhibitors include AWD-12-281(elbion); NCS-613 (INSERM); D-4418 (Chiroscience and Schering-Plough);CI-1018 or PD-168787 (Pfizer); benzodioxole compounds disclosed inWO99/16766 (Kyowa Hakko); K-34 (Kyowa Hakko); V-11294A (Napp);roflumilast (Byk-Gulden); pthalazinone compounds disclosed in WO99/47505(Byk-Gulden); Pumafentrine (Byk-Gulden, now Altana); arofylline(Almirall-Prodesfarma); VM554/UM565 (Vemalis); T-440 (Tanabe Seiyaku);and T2585 (Tanabe Seiyaku).

Representative muscarinic antagonists (i.e., anticholinergic agents)that can be used in combination with, and in addition to, the compoundsof this invention include, but are not limited to, atropine, atropinesulfate, atropine oxide, methylatropine nitrate, homatropinehydrobromide, hyoscyamine (d, l) hydrobromide, scopolamine hydrobromide,ipratropium bromide, oxitropium bromide, tiotropium bromide,methantheline, propantheline bromide, anisotropine methyl bromide,clidinium bromide, copyrrolate (Robinul), isopropamide iodide,mepenzolate bromide, tridihexethyl chloride (Pathilone), hexocycliummethylsulfate, cyclopentolate hydrochloride, tropicamide,trihexyphenidyl hydrochloride, pirenzepine, telenzepine, AF-DX 116 andmethoctramine and the like, or a pharmaceutically acceptable saltthereof; or, for those compounds listed as a salt, alternatepharmaceutically acceptable salt thereof.

Representative antihistamines (i.e., H₁-receptor antagonists) that canbe used in combination with the compounds of this invention include, butare not limited to, ethanolamines, such as carbinoxamine maleate,clemastine fumarate, diphenylhydramine hydrochloride and dimenhydrinate;ethylenediamines, such as pyrilamine amleate, tripelennaminehydrochloride and tripelennamine citrate; alkylamines, such aschlorpheniramine and acrivastine; piperazines, such as hydroxyzinehydrochloride, hydroxyzine pamoate, cyclizine hydrochloride, cyclizinelactate, meclizine hydrochloride and cetirizine hydrochloride;piperidines, such as astemizole, levocabastine hydrochloride, loratadineor its descarboethoxy analogue, terfenadine and fexofenadinehydrochloride; azelastine hydrochloride; and the like, or apharmaceutically acceptable salt thereof; or, for those compounds listedas a salt, alternate pharmaceutically acceptable salt thereof.

Suitable doses for the other therapeutic agents administered incombination with a compound of the invention are in the range of about0.05 μg/day to about 100 mg/day.

The following formulations illustrate representative pharmaceuticalcompositions of the present invention:

Formulation Example A

A dry powder for administration by inhalation is prepared as follows:

Ingredients Amount Compound of the invention 0.2 mg Lactose  25 mg

Representative Procedure: The compound of the invention is micronizedand then blended with lactose. This blended mixture is then loaded intoa gelatin inhalation cartridge. The contents of the cartridge areadministered using a powder inhaler.

Formulation Example B

A dry powder formulation for use in a dry powder inhalation device isprepared as follows:

Representative Procedure: A pharmaceutical composition is preparedhaving a bulk formulation ratio of micronized compound of the inventionto lactose of 1:200. The composition is packed into a dry powderinhalation device capable of delivering between about 10 μg and about100 μg of the compound of the invention per dose.

Formulation Example C

A dry powder for administration by inhalation in a metered dose inhaleris prepared as follows:

Representative Procedure: A suspension containing 5 wt. % of a compoundof the invention and 0.1 wt. % lecithin is prepared by dispersing 10 gof the compound of the invention as micronized particles with mean sizeless than 10 μm in a solution formed from 0.2 g of lecithin dissolved in200 mL of demineralized water. The suspension is spray dried and theresulting material is micronized to particles having a mean diameterless than 1.5 μm . The particles are loaded into cartridges withpressurized 1,1,1,2-tetrafluoroethane.

Formulation Example D

A pharmaceutical composition for use in a metered dose inhaler isprepared as follows:

Representative Procedure: A suspension containing 5 wt. % compound ofthe invention, 0.5 wt. % lecithin, and 0.5 wt. % trehalose is preparedby dispersing 5 g of active ingredient as micronized particles with meansize less than 10 μm in a colloidal solution formed from 0.5 g oftrehalose and 0.5 g of lecithin dissolved in 100 mL of demineralizedwater. The suspension is spray dried and the resulting material ismicronized to particles having a mean diameter less than 1.5 μm. Theparticles are loaded into canisters with pressurized1,1,1,2-tetrafluoroethane.

Formulation Example E

A pharmaceutical composition for use in a nebulizer inhaler is preparedas follows:

Representative Procedure: An aqueous aerosol formulation for use in anebulizer is prepared by dissolving 0.1 mg of the compound of theinvention in 1 mL of a 0.9% sodium chloride solution acidified withcitric acid. The mixture is stirred and sonicated until the activeingredient is dissolved. The pH of the solution is adjusted to a valuein the range of from 3 to 8 by the slow addition of NaOH.

Formulation Example F

Hard gelatin capsules for oral administration are prepared as follows:

Ingredients Amount Compound of the invention 250 mg Lactose(spray-dried) 200 mg Magnesium stearate  10 mg

Representative Procedure: The ingredients are thoroughly blended andthen loaded into a hard gelatin capsule (460 mg of composition percapsule).

Formulation Example G

A suspension for oral administration is prepared as follows:

Ingredients Amount Compound of the invention 1.0 g Fumaric acid 0.5 gSodium chloride 2.0 g Methyl paraben 0.15 g Propyl paraben 0.05 gGranulated sugar 25.5 g Sorbitol (70% solution) 12.85 g Veegum k(Vanderbilt Co.) 1.0 g Flavoring 0.035 mL Colorings 0.5 mg Distilledwater q.s. to 100 mL

Representative Procedure: The ingredients are mixed to form a suspensioncontaining 100 mg of active ingredient per 10 mL of suspension.

Formulation Example H

An injectable formulation is prepared as follows:

Ingredients Amount Compound of the invention 0.2 g Sodium acetate buffersolution (0.4M) 2.0 mL HCl (0.5N) or NaOH (0.5N) q.s. to pH 4 Water(distilled, sterile) q.s. to 20 mL

Representative Procedure: The above ingredients are blended and the pHis adjusted to 4±0.5 using 0.5 N HCl or 0.5 N NaOH.

Utility

The compounds of this invention possess both β₂ adrenergic receptoragonist and muscarinic receptor antagonist activity and therefore, suchcompounds are expected to be useful as therapeutic agents for treatingmedical conditions mediated by β₂ adrenergic receptors or muscarinicreceptors, i.e., medical conditions that are ameliorated by treatmentwith a β₂ adrenergic receptor agonist or a muscarinic receptorantagonist. See, for example, Eglen et al., Muscarinic ReceptorSubtypes: Pharmacology and Therapuetic Potential, DN&P 10(8), 462-469(1997); Emilien et al., Current Therapeutic Uses and Potential ofbeta-Adrenoceptor Agonists and Antagonists, European J Clinical Pharm.,53(6), 389-404 (1998); and references cited therein. Such medicalconditions include, by way of example, pulmonary disorders or diseasesassociated with reversible airway obstruction, such as chronicobstructive pulmonary disease (e.g., chronic and wheezy bronchitis andemphysema), asthma, pulmonary fibrosis and the like. Other conditionsinclude premature labor, depression, congestive heart failure, skindiseases (e.g., inflammatory, allergic, psoriatic and proliferative skindiseases), conditions where lowering peptic acidity is desirable (e.g.,peptic and gastric ulceration) and muscle wasting disease.

Accordingly, in one embodiment, this invention is directed to a methodfor treating a pulmonary disorder, the method comprising administeringto a patient in need of treatment a therapeutically effective amount ofa compound of formula I or a pharmaceutically acceptable salt or solvateor stereoisomer thereof. When used to treat a pulmonary disorder, thecompounds of this invention will typically be administered by inhalationin multiple doses per day, in a single daily dose or a single weeklydose. Generally, the dose for treating a pulmonary disorder will rangefrom about 10 μg/day to about 200 μg/day.

When administered by inhalation, the compounds of this inventiontypically have the effect of producing bronchodilation. Accordingly, inanother of its method aspects, this invention is directed to a method ofproducing bronchodilation in a patient, the method comprisingadministering to a patient requiring bronchodilation abronchodilation-producing amount of a compound of formula I or apharmaceutically acceptable salt or solvate or stereoisomer thereof.Generally, the dose for producing bronchodilation will range from about10 μg/day to about 200 μg/day.

In one embodiment, this invention is directed to a method of treatingchronic obstructive pulmonary disease or asthma, the method comprisingadministering to a patient in need of treatment a therapeuticallyeffective amount of a compound of formula I or a pharmaceuticallyacceptable salt or solvate or stereoisomer thereof. When used to treat aCOPD or asthma, the compounds of this invention will typically beadministered by inhalation in multiple doses per day or in a singledaily dose. Generally, the dose for treating COPD or asthma will rangefrom about 10 μg/day to about 200 μg/day.

As used herein, COPD includes chronic obstructive bronchitis andemphysema (see, for example, Barnes, Chronic Obstructive PulmonaryDisease, N Engl J Med 2000: 343:269-78).

When used to treat a pulmonary disorder, the compounds of this inventionare optionally administered in combination with other therapeuticagents. In particular, by combining the compounds of this invention witha steroidal anti-inflammatory agent (e.g. a corticosteroid), thepharmaceutical compositions of this invention can provide tripletherapy, i.e., β₂ adrenergic receptor agonist, muscarinic receptorantagonist and anti-inflammatory activity, using only two activecomponents. Since pharmaceutical compositions containing two activecomponents are typically easier to formulate compared to compositionscontaining three active components, such two component compositionsprovide a significant advantage over compositions containing threeactive components. Accordingly, in a particular embodiment, thepharmaceutical compositions and methods of this invention furthercomprise a therapeutically effective amount of a steroidalanti-inflammatory agent.

Compounds of this invention exhibit both muscarinic receptor antagonistand β₂ adrenergic receptor agonist activity. Accordingly, among otherproperties, compounds of particular interest are those that demonstratean inhibitory constant K_(i) value for binding at the M₃ muscarinicreceptor and an EC₅₀ value for β₂ adrenergic receptor agonist activityof less than about 100 nM; particularly less than 10 nM. Among thesecompounds, compounds of special interest include those having muscarinicactivity, expressed in terms of the inhibitory constant K_(i) forbinding at the M₃ muscarinic receptor, that is about equal to thecompound's β₂ adrenergic agonist activity, expressed in terms of thehalf maximal effective concentration EC₅₀, as determined in the in vitroassays described herein, or in similar assays. For example, compounds ofparticular interest are those having a ratio of the inhibitory constantK_(i) for the M₃ muscarinic receptor to the EC₅₀ for the β₂ adrenergicreceptor ranging from about 30:1 to about 1:30; including about 20:1 toabout 1:20; such as about 10:1 to about 1:10.

Since compounds of this invention possess both β₂ adrenergic agonistactivity and muscarinic receptor antagonist activity, such compounds arealso useful as research tools for investigating or studying biologicalsystems or samples having β₂ adrenergic receptors or muscarinicreceptors, or for discovering new compounds having both β₂ adrenergicagonist activity and muscarinic receptor antagonist activity. Suchbiological systems or samples may comprise β₂ adrenergic receptorsand/or muscarinic receptors. Any suitable biological system or samplehaving β₂ adrenergic and/or muscarinic receptors may be employed in suchstudies which may be conducted either in vitro or in vivo.Representative biological systems or samples suitable for such studiesinclude, but are not limited to, cells, cellular extracts, plasmamembranes, tissue samples, mammals (such as mice, rats, guinea pigs,rabbits, dogs, pigs, etc.), and the like.

In this embodiment, a biological system or sample comprising a β₂adrenergic receptor or a muscarinic receptor is contacted with a , 2adrenergic receptor-agonizing or muscarinic receptor-antagonizing amountof a compound of this invention. The effects are then determined usingconventional procedures and equipment, such as radioligand bindingassays and functional assays. Such functional assays includeligand-mediated changes in intracellular cyclic adenosine monophosphate(cAMP), ligand-mediated changes in activity of the enzyme adenylylcyclase (which synthesizes cAMP), ligand-mediated changes inincorporation of guanosine 5′-O-(-thio)triphosphate ([³⁵S]GTP S) intoisolated membranes via receptor catalyzed exchange of [³⁵S]GTP S forGDP, ligand-mediated changes in free intracellular calcium ions(measured, for example, with a fluorescence-linked imaging plate readeror FLIPR® from Molecular Devices, Inc.). A compound of this inventionwill agonize or cause activation of a β₂ adrenergic receptor andantagonize or decrease the activation of muscarinic receptors in any ofthe functional assays listed above, or assays of a similar nature. Theamount of compound used in these studies will typically range from about0.1 nanomolar to about 100 nanomolar.

Additionally, the compounds of this invention can be used as researchtools for discovering new compounds that have both a β₂ adrenergicreceptor agonist and muscarinic receptor antagonist activity. In thisembodiment, a β₂ adrenergic receptor and muscarinic receptor bindingdata (for example, as determined by in vitro radioligand displacementassays) for a test compound or a group of test compounds is compared tothe β₂ adrenergic receptor and muscarinic receptor binding data for acompound of this invention to identify those test compounds that haveabout equal or superior β₂ adrenergic receptor and/or muscarinicreceptor binding, if any. This aspect of the invention includes, asseparate embodiments, both the generation of comparison data (using theappropriate assays) and the analysis of the test data to identify testcompounds of interest.

In some cases, compounds of this invention may possess either weakmuscarinic receptor antagonist activity or weak β₂ adrenergic receptoragonist activity. In these cases, those of ordinary skill in the artwill recognize that such compounds still have utility as primarilyeither a β₂ adrenergic receptor agonist or a muscarinic receptorantagonist, respectively.

The properties and utility of the compounds of this invention can bedemonstrated using various in vitro and in vivo assays well-known tothose skilled in the art. For example, representative assays aredescribed in further detail in the following Examples.

EXAMPLES

The following Preparations and Examples are provided to illustratespecific embodiments of this invention. These specific embodiments,however, are not intended to limit the scope of this invention in anyway unless specifically indicated.

The following abbreviations have the following meanings unless otherwiseindicated and any other abbreviations used herein and not defined havetheir standard meaning:

AC adenylyl cyclase Ach acetylcholine ATCC American Type CultureCollection BSA bovine serum albumin cAMP 3′-5′ cyclic adenosinemonophosphate CHO Chinese hamster ovary cM₅ cloned chimpanzee M₅receptor DCM dichloromethane (i.e., methylene chloride) DIPEAN,N-diisopropylethylamine dPBS Dulbecco's phosphate buffered saline DMEMDulbecco's Modified Eagle's Medium DMSO dimethyl sulfoxide EDTAethylenediaminetetraacetic acid Emax maximal efficacy EtOAc ethylacetate EtOH ethanol FBS fetal bovine serum FLIPR fluorometric imagingplate reader Gly glycine HATUO-(7-azabenzotriazol-1-yl-N,N,N′,N′-tetramethyluroniumhexafluorophosphate HBSS Hank's buffered salt solution HEK humanembryonic kidney cells HEPES4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid hM₁ cloned human M₁receptor hM₂ cloned human M₂ receptor hM₃ cloned human M₃ receptor hM₄cloned human M₄ receptor hM₅ cloned human M₅ receptor HPLChigh-performance liquid chromatography IBMX 3-isobutyl-1-methylxanthine% Eff % efficacy PBS phosphate buffered saline PyBOPbenzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate rpmrotations per minute TFA trifluoroacetic acid THF tetrahydrofuran Tristris(hydroxymethyl)aminomethane

Unless noted otherwise, reagents, starting materials and solvents werepurchased from commercial suppliers (such as Aldrich, Fluka, Sigma andthe like) and were used without further purification.

In the examples described below, HPLC analysis was conducted using anAgilent (Palo Alto, Calif.) Series 1100 instrument with Zorbax Bonus RP2.1×50 mm columns, supplied by Agilent, (a C14 column), having a 3.5micron particle size. Detection was by UV absorbance at 214 nm. HPLC10-70 data was obtained with a flow rate of 0.5 mL/minute of 10%-70% Bover 6 minutes. Mobile phase A was 2% -98% -0.1% ACN-H₂O-TFA; and mobilephase B was 90% -10% -0.1% ACN-H₂O-TFA. Using the mobile phases A and Bdescribed above, HPLC 5-35 data and HPLC 10-90 data were obtained with a5 minute gradient.

Liquid chromatography mass spectrometry (LCMS) data were obtained withan Applied Biosystems (Foster City, Calif.) model API-150EX instrument.LCMS 10-90 data was obtained with a 10% -90% mobile phase B over a 5minute gradient.

Small scale purification was conducted using an API 150EX PrepWorkstation system from Applied Biosystems. The mobile phase was A:water+0.05% v/v TFA; and B: acetonitrile+0.05% v/v TFA. For arrays(typically about 3 to 50 mg recovered sample size) the followingconditions were used: 20 mL/min flow rate; 15 min gradients and a 20mm×50 mm Prism RP column with 5 micron particles (ThermoHypersil-Keystone, Bellefonte, Pa.). For larger scale purifications(typically greater than 100 mg crude sample), the following conditionswere used: 60 mL/min flow rate; 30 min gradients and a 41.4 mm×250 mmMicrosorb BDS column with 10 micron particles (Varian, Palo Alto,Calif.).

The specific rotation for chiral compounds (indicated as [α]²⁰D) wasmeasured using a Jasco Polarimeter (Model P-1010) with a tungstenhalogen light source and a 589 nm filter at 20° C. Samples of testcompounds were typically measured at 1 mg/mL water.

Preparation 1 8-Benzyloxy-5-(2-bromoacetyl)-1H-quinolin-2-one (a)8-Acetoxy-1H-quinolin-2-one

8-Hydroxyquinoline-N-oxide (160.0 g, 1.0 mol), commercially-availablefrom Aldrich, Milwaukee, Wis., and acetic anhydride (800 mL, 8.4 mol)were heated at 100° C. for 3 h and then cooled in ice. The product wascollected on a Buchner funnel, washed with acetic anhydride (2×100 mL)and dried under reduced pressure to give 8-acetoxy-1H-quinolin-2-one(144 g) as a solid.

(b) 5-Acetyl-8-hydroxy-1H-quinolin-2-one

A slurry of aluminum chloride (85.7 g, 640 mmol) in 1,2-dichloroethane(280 mL) was cooled in ice, and the product from step (a) (56.8 g, 280mmol) was added. The mixture was warmed to room temperature and thenheated at 85° C. After 30 min, acetyl chloride (1.5 mL, 21 mmol) wasadded and the mixture was heated an additional 60 min. The reactionmixture was then cooled and added to IN hydrochloric acid (3 L) at 0° C.with good stirring. After stirring for 2 h, the solids were collected ona Buchner funnel, washed with water (3×250 mL) and dried under reducedpressure. The crude product isolated from several batches (135 g) wascombined and triturated with dichloromethane (4 L) for 6 h. Theresulting solid was collected on a Buchner funnel and dried underreduced pressure to give the title compound (121 g).

(c) 5-Acetyl-8-benzyloxy-1H-quinolin-2-one

To the product from step (b) (37.7 g, 186 mmol) was addedN,N-dimethylformamide (200 mL) and potassium carbonate (34.5 g, 250mmol) followed by benzyl bromide (31.8 g, 186 mmol). The mixture wasstirred at room temperature for 2.25 hour and then poured into saturatedsodium chloride (3.5 L) at 0° C. and stirred for 1 hour. The product wascollected and dried on a Buchner funnel for 1 hour, and the resultingsolids were dissolved in dichloromethane (2 L) and this mixture wasdried over sodium sulfate. The solution was filtered through a pad ofCelite which was then washed with dichloromethane (5×200 mL). Thecombined filtrate was then concentrated to dryness and the resultingsolids were triturated with ether (500 mL) for 2 h. The product wascollected on a Buchner funnel, washed with ether (2×250 mL) and driedunder reduced pressure to give the title compound (44 g) as a powder.

(d) 8-Benzyloxy-5-(2-bromoacetyl)-1H-quinolin-2-one

The product from step (c) (20.0 g, 68.2 mmol) was dissolved indichloromethane (200 mL) and cooled to 0° C. Boron trifluoride diethyletherate (10.4 mL, 82.0 mmol) was added via syringe and the mixture waswarmed to room temperature to give a thick suspension. The suspensionwas heated at 45° C. (oil bath) and a solution of bromine (11.5 g, 72.0mmol) in dichloromethane (100 mL) was added over 40 min. The mixture waskept at 45° C. for an additional 15 min and then cooled to roomtemperature. The mixture was concentrated under reduced pressure andthen triturated with 10% aqueous sodium carbonate (200 mL) for 1 hour.The solids were collected on a Buchner funnel, washed with water (4×100mL) and dried under reduced pressure. The product of two runs wascombined for purification. The crude product (52 g) was triturated with50% methanol in chloroform (500 mL) for 1 hour. The product wascollected on a Buchner funnel and washed with 50% methanol in chloroform(2×50 mL) and methanol (2×50 mL). The solid was dried under reducedpressure to give the title compound (34.1 g) as a powder.

Preparation 28-Benzyloxy-5-[(R)-2-bromo-1-(tert-butyldimethylsflanyloxy)ethyl]-1H-quinolin-2-one(a) 8-Benzyloxy-5-((R)-2-bromo-1-hydroxyethyl)-1H-quinolin-2-one

(R)-(+)-α,α-Diphenylprolinol (30.0 g, 117 mmol) and trimethylboroxine(11.1 mL, 78 mmol) were combined in toluene (300 mL) and stirred at roomtemperature for 30 min. The mixture was placed in a 150° C. oil bath andliquid was distilled off. Toluene was added in 20 mL aliquots anddistillation was continued for 4 h. A total of 300 mL toluene was added.The mixture was then cooled to room temperature. A 500 μL aliquot wasevaporated to dryness and weighed (246 mg) to determine that theconcentration of catalyst was 1.8 M.

8-Benzyloxy 5-(2-bromoacetyl)-1H-quinolin-2-one (90.0 g, 243 mmol) wasplaced under nitrogen and tetrahydrofuran (900 mL) was added followed bythe catalyst described above (1.8 M in toluene, 15 mL, 27 mmol). Thesuspension was cooled to −10±5° C. in an ice/isopropanol bath. Borane(1.0 M in THF, 294 mL, 294 mmol) was added over 4 h. The reaction wasthen stirred an additional 45 min at −10° C. and then methanol (250 mL)was added slowly. The mixture was concentrated under vacuum and theresidue was dissolved in boiling acetonitrile (1.3 L), filtered whilehot and then cooled to room temperature. The crystals were filtered,washed with acetonitrile and dried under vacuum to give the titlecompound (72.5 g, 196 mmol, 81% yield, 95% ee, 95% pure by HPLC).

(b)8-Benzyloxy-5-[(R)-2-bromo-1-(tert-butyldimethylsilanyloxy)ethyl]-1H-quinolin-2-one

To the product of step (b) (70.2 g, 189 mmol) was addedN,N-dimethylformamide (260 mL) and this mixture was cooled in an icebath under nitrogen. 2,6-Lutidine (40.3 g, 376 mmol) was added over 5min and then tert-butyldimethylsilyl trifluoromethanesulfonate (99.8 g,378 mmol) was added slowly while maintaining the temperature below 20°C. The mixture was allowed to warm to room temperature for 45 min.Methanol (45 mL) was added to the mixture dropwise over 10 min and themixture was partitioned between ethyl acetate/cyclohexane(1:1, 500 mL)and water/brine (1:1, 500 mL). The organics were washed twice more withwater/brine (1:1, 500 mL each). The combined organics were evaporatedunder reduced pressure to give a light yellow oil. Two separate portionsof cyclohexane (400 mL) were added to the oil and distillation continueduntil a thick white slurry was formed. Cyclohexane (300 mL) was added tothe slurry and the resulting white crystals were filtered, washed withcyclohexane (300 mL) and dried under reduced pressure to give the titlecompound (75.4 g, 151 mmol, 80% yield, 98.6% ee).

Preparation 3A8-Benzyloxy-5-[(R)-2-(N-benzylamino)-1-(tert-butyldimethylsilanyloxy)ethyl]-1H-quinolin-2-one

A stirred solution of the product of Preparation 2 (1.00 g, 2.05 mmol)and benzylamine (493 μL, 4.51 mmol) in DMSO (1.7 mL) was heated at 105°C. for 4 h. The reaction mixture was allowed to cool and was thendiluted with EtOAc (10 mL) and the organic layer was washed withsaturated aqueous ammonium chloride solution (5 mL) and 1N sodiumhydroxide (5 mL), dried (MgSO₄) and solvent removed under reducedpressure. The crude residue was purified by column chromatography (50%EtOAc/hexanes) to give the title compound (700 mg, 67%). MS m/z: [M+H⁺]calcd for C₃₁H₃₈N₂O₃Si 515.27; found 515.5.

Preparation 3B8-Benzyloxy-5-[(R)-2-(N-benzylamino)-1-(tert-butyldimethylsilanyloxy)ethyl]-1H-quinolin-2-one

To a 500 mL three-necked round-bottom flask was added8-benzyloxy-5-[(R)-2-bromo-1-(tert-butyldimethylsilanyloxy)ethyl]-1H-quinolin-2-one(43 g, 0.124 mol, about 95% chiral purity), 1-methyl-2-pyrrolidinone(210 mL) and benzylamine (28.3 g, 0.37 mol). The resulting mixture wasflushed with nitrogen and then stirred at 90° C. for 6 hours. Themixture was then cooled to room temperature and water (300 mL) and ethylacetate (300 mL) were added. The layers were separated and the organiclayer was washed with water (200 mL), a 1:1 mixture of water and aqueoussaturated sodium chloride solution (200 mL), and water (200 mL). Theorganic layer was then dried over magnesium sulfate, filtered andconcentrated under reduced pressure to provide the title compound as anorange oil.

To the orange oil was added heptane (200 mL) and ethyl acetate (200 mL)and the resulting mixture was heated to 65° C. to produce a clearsolution. This solution was cooled to room temperature and allowed tostand overnight (about 16 hours) at which time a precipitate had formed.The precipitate was collected by filtration to givestereochemically-impure title compound (8.85 g, 79.6% ee). The filtratewas concentrated under reduced pressure to give the title compound (38.6g, 99.4% ee). This material was combined with a previous batch ofmaterial (19.2 g, 99.5% ee) and heptane (250 mL) and ethyl acetate (100mL) were added. This mixture was heated to 80° C. (hazy to clearsolution) and then cooled to room temperature and allowed to standovernight. The resulting precipitate was collected by filtration toafford the title compound as a white solid (36.8 g. 98.4% ee, 99.9%chemical purity). The filtrate was concentrated under reduced pressureand the residue was dissolved in heptane (100 mL). The resulting solidswere collected to give the title compound as a tan solid (24 g, 100%chiral purity, 95% ee).

Preparation 45-[(R)-2-Amino-1-(tert-butyldimethylsilanyloxy)ethyl]-8-hydroxy-1H-quinolin-2-oneAcetic Acid Salt

8-Benzyloxy-5-[(R)-2-(N-benzylamino)-1-(tert-butyldimethylsilanyloxy)ethyl]-1H-quinolin-2-one(100 g, 194 mmol) and acetic acid (17.5 mL, 291 mmol) were dissolved inmethanol (1 L). The clear solution was purged with nitrogen and thenpalladium hydroxide on carbon (20 g, 20 wt. % Pd (dry basis), wet (about50% water)) was added. Hydrogen gas was bubbled through the stirredsolution at room temperature for 6 hours during which time a thickslurry developed. The reaction mixture was then purged with nitrogen andmethanol (1 L) was added. The resulting mixture was stirred for about 30minutes (to dissolve the product) and then the mixture was filteredthrough a pad of Celite. The filtrate was concentrated under reducedpressure to a volume of about 500 mL and, to the resulting slurry, wasadded ethanol (500 mL). The resulting mixture was again concentratedunder reduced pressure to a volume of about 500 mL and the resultingprecipitate was collected by filtration and dried to provide the titlecompound as a yellow-white solid (65 g, 85% yield, >98% purity).

Preparation 5 Ethyl 3-(N-Benzyl-N-tert-butoxycarbonylamino)propionate

Ethyl 3-benzylaminopropionate (10 g, 48.2 mmol) was dissolved indichloromethane (100 mL) and to this solution was added triethylamine(6.72 mL, 53 mmol). This mixture was flushed with dry nitrogen anddi-tert-butyl dicarbonate (11 g, 51 mmol) was added. The resultingmixture was stirred at room temperature for 3 hours. The reaction wasthen concentrated under reduced pressure and the residue was dissolvedin ethyl acetate. This solution was washed with aqueous hydrochloricacid (1N,×2), aqueous sodium hydroxide (1N,×2) and brine (×1). Theorganic layer was then dried over magnesium sulfate, filtered andconcentrated under reduced pressure to afford the title compound as aclear colorless oil (15.6 g). ¹H NMR (CDCl₃, δ) 7.4-7.2 (m, SH), 4.45(br s, 2H), 4.1 (q, 2H), 3.5 (m, 2H), 2.55 (m, 2H), 1.5 (m, 9H), 1.25(t, 3H).

Preparation 6 4-(N-Benzyl-N-tert-butoxycarbonylamino)-2-methylbutan-2-ol

Under nitrogen, THF (30 mL) and methyllithium (1.6 M in diethyl ether,35 mL) were added via a cannula to a dry 500 mL round-bottomed flaskequipped with a stir bar and a 60 mL graduated dropping funnel. Thecannula was then rinsed with a further portion of THF (10 mL). The flaskwas cooled to −78° C. and a solution of the product of Preparation 5(6.16 g, 20 mmol) in THF (30 mL) was added dropwise from the droppingfunnel over a period of 25 minutes. The resulting reaction mixture wasstirred at −78° C. for 1.5 hours and then quenched with water. Thismixture was warmed to room temperature and diluted with ethyl acetate.The resulting mixture was stirred at room temperature overnight. Themixture was then transferred to a separatory funnel and the layers wereseparated. The organic layer was washed with saturated aqueous sodiumbicarbonate solution (×2) and brine (×1). The organic layer was thendried over magnesium sulfate, filtered and concentrated under reducedpressure to afford the title compound as a clear colorless oil (6 g).LCMS m/z: 294.3 (MH⁺).

Preparation 7 4-(Benzylmethylamino)-2-methylbutan-2-ol

The product of Preparation 6 (2.4 g, 8.2 mmol) was dissolved indichloromethane (5 mL) and trifluoroacetic acid (5 mL) was added. Theresulting mixture was stirred at room temperature for 2 hours. Thesolvent was evaporated under reduced pressure and the residue wasdissolved in dichloromethane. The solution was transferred into aseparatory funnel and washed with aqueous sodium hydroxide solution(1N,×3) and brine (×1). The organic layer was then dried over magnesiumsulfate, filtered and concentrated under reduced pressure.

The resulting oil was dissolved in methanol (10 mL) and to this solutionwas added paraformaldehyde (2.5 g, 82 mmol). This mixture was heated atreflux for 3 hours and then cooled to room temperature. Sodiumtriacetoxyborohydride (5.2 g, 24.8 mmol) was added and the reactionmixture was stirred at room temperature overnight. The methanol wasevaporated under reduced pressure and the residue was dissolved indichloromethane. This solution was washed with aqueous sodium hydroxidesolution (1N,×3) and brine (×1). The organic layer was then dried overmagnesium sulfate, filtered and concentrated under reduced pressure. Theresidue was purified by silica gel column chromatography eluting with10% methanol/chloroform to give the title compound as a clear paleyellow oil (1.49 g). ¹H NMR (CDCl₃, δ): 7.3 (m, SH), 3.5 (s, 2H), 2.65(m, 2H), 2.2 (s, 2H), 1.65 (m, 2H), 1.1 (s, 6H).

Preparation 8 Biphenyl-2-ylcarbamic Acid3-(Benzylmethylamino)-1,1-dimethylpropyl Ester

The product of Preparation 7 (500 mg, 2.4 mmol) was dissolved inN,N-dimethylformamide (1.3 mL) and to this solution was addedbiphenylisocyanate (455 μL, 2.65 mmol). The resulting mixture was heatedat 70° C. overnight and then ethyl acetate was added. This mixture waswashed with aqueous sodium hydroxide solution (1N,×2) and brine (×1).The organic layer was then dried over magnesium sulfate, filtered andconcentrated under reduced pressure to provide the title compound as anoil. LCMS m/z: 403.5 (MH⁺).

Preparation 9 Biphenyl-2-ylcarbamic Acid3-[(9-Hydroxynonyl)methylamino]-1,1-dimethylpropyl Ester

The product of Preparation 8 (970 mg, 2.4 mmol) was dissolved in ethanol(10 mL) and nitrogen gas was bubbled through this solution for 5minutes. Palladium on carbon (10%, degussa type, 1 g) was added and thereaction mixture was stirred at room temperature overnight under ahydrogen-filled balloon. Nitrogen gas was then bubbled through thereaction mixture. The mixture was then filtered through a pad of Celiteand the Celite pad was washed with ethanol. The filtrate wasconcentrated under reduced pressure.

The residue was dissolved in acetonitrile (10 mL) and to this solutionwas added 9-bromo-1-nonanol (642 mg, 2.88 mmol) anddiisopropylethylamine (627 μL, 3.6 mmol). The reaction mixture washeated at 60° C. overnight and then cooled to room temperature andconcentrated under reduced pressure. The residue was dissolved indichloromethane and the resulting solution was washed with aqueoussodium dihydrogenphosphate solution (1M,×2), dried over magnesiumsulfate, filtered and concentrated under reduced pressure. The resultingoil was then chromatographed over silica gel eluting with 5%methanol/chloroform/0.5% NH₄OH to afford the title compound (284 mg).LCMS m/z: 455.3 (MH⁺).

EXAMPLE 1 Biphenyl-2-ylcarbamic Acid3-({9-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydro-quinolin-5-yl)ethylamino]nonyl}methylamino)-1,1-dimethylpropylEster

The product of Preparation 9 (284 mg, 0.63 mmol) was dissolved indichloromethane (5 mL) to this solution was added diisopropylethylamine(330 μL, 1.88 mmol) and dimethyl sulfoxide (135 μL, 1.88 mmol). Theflask was sealed under a nitrogen atmosphere and cooled to −15° C.Pyridine sulfur trioxide complex (300 mg, 1.88 mmol) was added and thereaction mixture was stirred at −15° C. for 1 h. The reaction was thenquenched by adding water and the layers were separated. The organiclayer was washed with water (×3), aqueous saturated sodium bicarbonatesolution (×1) and brine (×1). The organic layer was then dried overmagnesium sulfate, filtered and concentrated under reduced pressure.

The residue was dissolved in a 1:1 mixture of dichloromethane (1 mL) andmethanol (1 mL) and to this solution was added the product ofPreparation 4 (316 mg, 0.94 mmol) and sodium triacetoxyborohydride (400mg, 1.88 mmol). The reaction mixture was stirred at room temperatureovernight and then concentrated under reduced pressure.

The residue was dissolved in dichloromethane (5 mL) and to this solutionwas added triethylamine trihydrofluoride (1.03 mL, 6.3 mmol). Thereaction mixture was stirred at room temperature overnight and then thesolvent was evaporated under reduced pressure. The resulting foam wasdissolved in a 1:1 mixture of acetic acid (3 mL) and water (3 mL) andthis mixture was purified by reverse phase preparative HPLC to affordthe title compound as a white lyophilized solid (18.0 mg). LCMS m/z:657.5 (MH⁺).

Preparation 10 (2-Bromophenyl)carbamic Acid3-(Benzylmethylamino)-1,1-dimethylpropyl Ester

Following the procedure of Preparation 8 and using2-bromophenylisocyanate in place of biphenylisocyanate, the titlecompound is prepared.

Preparation 11 (2-Thien-3-ylphenyl)carbamic Acid3-(Benzylmethylamino)-1,1-dimethylpropyl Ester

The product of Preparation 10 (0.41 mmol) is dissolved in a 1:1 mixtureof ethylene glycol dimethyl ether and N,N-dimethylformamide (2 mL total)and to this solution is added 3-thiopheneboronic acid (80 mg, 0.62 mmol)and aqueous potassium carbonate solution (4M, 205 μL, 0.82 mmol). Thevial is flushed with nitrogen and then[1,1′-bis(diphenylphosphino)ferrocene]dicloropalladium(II) (catalyticamount) is added and the vial is sealed and heated on a shaker block at86° C. overnight. The solvent is removed under reduced pressure and theresidue is dissolved in dichloromethane and washed with aqueous sodiumbisulfate (1M×2), aqueous sodium hydroxide (1N×2), brine (×1) and theorganic layer is dried over magnesium sulfate, filtered and concentratedunder reduced pressure. The residue is purified by silica gelchromatography to provide the title compound.

EXAMPLE 2 (2-Thien-3-ylphenyl)carbamic Acid3-({9-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}methylamino)-1,1-dimethylpropylEster

Using the product of Preparation 11 and the procedures of Preparation 9and Example 1, the title is prepared.

Preparation 12 Biphenyl-2-ylcarbamic Acid3-[(2-tert-Butoxycarbonylaminoethyl)methylamino]-1,1-dimethylpropylEster

To a stirred solution of the debenzylated product of Preparation 9 (6.76mmol) and DIPEA (3.54 mL, 20.3 mmol) in acetonitrile (67.6 mL) at 50° C.is added 2-tert-butoxycarbonylaminoethyl bromide (1.82 g, 8.11 mmol) andthe reaction mixture is heated at 50° C. overnight. The solvent is thenremoved under reduced pressure and the residue is purified by flashchromatography to yield the title compound.

Preparation 13 Biphenyl-2-ylcarbamic Acid3-[(2-Aminoethyl)methylamino]-1,1-dimethylpropyl Ester

The product of Preparation 12 is dissolved in TFA/DCM (25%, 52 mL) andstirred at room temperature for 2 h. The solvent is then removed underreduced pressure and the residue is purified by flash chromatography toyield the title compound.

Preparation 14 Biphenyl-2-ylcarbamic Acid3-{[2-(4-Aminomethylbenzoylamino)ethyl]methyl-amino}-1,1-dimethylpropylEster

To a stirred solution of the product of Preparation 13 (1 mmol),4-(tert-butoxycarbonylaminomethyl)benzoic acid (301 mg, 1.2 mmol) andHATU (456 mg, 1.2 mmol) in DMF (2 mL) is added DIPEA (226 μL, 1.3 mmol).The reaction mixture is stirred at room temperature overnight and thenthe solvent is removed under reduced pressure. The resulting residue isdissolved in TFA/DCM (25%, 10 mL) and this mixture is stirred at roomtemperature for 2 h. The solvent is removed under reduced pressure andthe residue is purified by flash chromatography to yield the titlecompound.

Preparation 15 Biphenyl-2-ylcarbamic Acid3-{[2-(4-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydro-quinolin-5-yl)-2-(tert-butyldimethylsilanyloxy)ethylamino]methyl}benzoylamino)-ethyl]methylamino}-1,1-dimethylpropylEster

A solution of the product of Preparation 14 (1.1 mmol), the product ofPreparation 2 (634 mg, 1.3 mmol), sodium bicarbonate (277 mg, 3.3 mmol)and sodium iodide (215 mg, 1.43 mmol) in THF (0.55 mL) is heated at 80°C. for 12 h. The solvent is removed under reduced pressure and the cruderesidue is purified by flash chromatography to give the title compound.

Preparation 16 Biphenyl-2-ylcarbamic Acid3-{[2-(4-{[(R)-2-(8-Benzyloxy-2-oxo-1,2-dihydro-quinolin-5-yl)-2-hydroxyethylamino]methyl}benzoylamino)ethyl]methylamino}-1,1-dimethylpropylester

To a stirred solution of the product of Preparation 15 (0.36 mmol) indichloromethane (3.6 mL) is added triethylamine trihydrofluoride (117μL, 0.72 mmol). The reaction mixture is stirred for 10 h and is thendiluted with dichloromethane (10 mL) and washed with saturated aqueoussodium bicarbonate solution (5 mL). The organic phase is dried (MgSO₄)and the solvent is removed under reduced pressure to give the titlecompound.

EXAMPLE 3 Biphenyl-2-ylcarbamic Acid3-{[2-(4-{[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzoylamino)ethyl]methylamino}-1,1-dimethylpropylEster

To a stirred solution of the product of Preparation 16 (0.36 mmol) inethanol (3.6 mL) is added palladium (10 wt. % (dry basis) on activatedcarbon) (275 mg) and the reaction mixture is placed under a hydrogenatmosphere and stirred overnight. The reaction mixture is then filteredand the solvent is removed under reduced pressure. The crude residue ispurified by preparative HPLC to yield the title compound as theditrifluoroacetate salt.

Preparation 17 Methyl 4-Iodophenylacetate

To a stirred solution of 4-iodophenylacetic acid (5.0 g, 19.1 mmol) inMeOH (200 mL) was added 4N hydrochloric acid in dioxane (10 mL). Thereaction mixture was stirred for 24 h at room temperature and then thesolvent was removed under reduced pressure to give the title compound(5.17 g, 98% yield), which was used without further purification.

Preparation 18 Methyl [4-(4-Hydroxybut-1-ynyl)phenyl]acetate

To a stirred solution of the product of Preparation 17 (4.5 g, 16.3mmol) in diethylamine (100 mL) was added but-3-yn-1-ol (1.9 mL, 32.6mmol), Pd(PPh₃)₂Cl₂ (500 mg, 1.63 mmol) and CuI (154 mg, 0.815 mmol) andresulting mixture was stirred for 17 h at room temperature. The solventwas then removed under reduced pressure and the residue was dissolved indiethyl ether (200 mL) and this solution was filtered to remove salts.The solvent was then removed under reduced pressure and the crudeproduct was purified by silica gel chromatography (60% EtOAc/Hexane) toafford the title intermediate (3.03 g, 91% yield).

Preparation 19 Methyl [4-(4-Hydroxybutyl)phenyl]acetate

A stirred solution of the product of Preparation 18 (2.8 g, 12.8 mmol)in methanol (50 mL) was flushed with nitrogen and then 10% palladium oncarbon (400 mg, 20% wt/wt) was added. The reaction flask was thenalternately placed under vacuum and flushed with hydrogen for cycles andthen stirred under hydrogen for 14 h. The reaction mixture was flushedwith nitrogen and then filtered and the solvent removed under reducedpressure to give the title compound (2.75 g, 97% yield), which was usedwithout further purification.

Preparation 20 Methyl[4-(4-{[3-(Biphenyl-2-ylcarbamoyloxy)-3-methylbutyl]methylamino}butyl)-phenyl]acetate(a) Methyl {4-[4-(Toluene-4-sulfonyloxy)butyl]phenyl}acetate

To a stirred solution of the product of Preparation 19 (2.6 g, 12.5mmol) in THF (100 mL) was added DABCO (2.6 g, 25.0 mmol) and thenp-toluenesulfonyl chloride (2.44 g, 13.75 mmol). The reaction mixturewas stirred at room temperature for 23 h and then solvent was removedunder reduced pressure and the residue was dissolved in dichloromethane(200 mL). The organic layer was then washed with water (2×100 mL), 1Nhydrochloric acid (100 mL), aqueous saturated sodium chloride solution(100 mL), dried (MgSO₄), filtered and the solvent removed under reducedpressure to give the title compound, which was used without furtherpurification.

(b) Methyl[4-(4-{[3-(Biphenyl-2-ylcarbamoyloxy)-3-methylbutyl]methylamino}butyl)-phenyl]acetate

To the crude product from step (a) is added DMF (50 mL),diisopropylethylamine (3.0 mL, 17.3 mmol) and the debenzylated productof Preparation 9 (8.1 mmol). The reaction mixture is stirred at roomtemperature for 18 h and then the solvent is removed under reducedpressure to give the title compound.

Preparation 21 Biphenyl-2-ylcarbamic Acid3-({4-[4-(2-Hydroxyethyl)phenyl]butyl}methylamino)-1,1-dimethylpropylEster

To a stirred solution of the product of Preparation 20 (4.0 mmol) in THF(100 mL) is added dropwise DIBAL-H (24 mL, 24 mmol, 1.0 M in THF). Afterthe addition is complete, the reaction mixture is stirred for 3 h andthen quenched by slow addition of methanol (until gas evolution ceased).The mixture is then concentrated to dryness and the residue is purifiedby flash chromatography to give the title compound.

EXAMPLE 4 Biphenyl-2-ylcarbamic Acid3-{[4-(4-{2-[(R)-2-Hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenyl)butyl]methylamino}-1,1-dimethyl-propylEster

Using the product of Preparation 21 and the procedures of Example 1, thetitle compound is prepared.

Preparation 22 Methyl 4-Amino-5-chloro-2-methoxybenzoate

To a solution of 4-amino-5-chloro-3-methoxybenzoic acid (1.008 g, 5.0mmol) in a mixture of toluene (9 mL) and methanol (1 mL) at 0° C. wasadded (trimethylsilyl)diazomethane (2.0 M in hexane, 3.0 mL, 6.0 mmol)dropwise. The reaction mixture was then warmed to room temperature andstirred for 16 h. Excess (trimethylsilyl)diazomethane was quenched byadding acetic acid until the bright yellow color of the reaction mixturedisappeared. The mixture was then concentrated in vacuo to give thetitle compound as an off-white solid, which was used without furtherpurification.

Preparation 23 Methyl 4-Acryloylamino-5-chloro-2-methoxybenzoate

To crude product of Preparation 22 was added dichloromethane (10 mL, 0.5M) and triethylamine (2.1 mL, 15 mmol). This mixture was cooled to 0° C.and acryloyl chloride (812 μL, 10 mmol) was added dropwise withstirring. After 2 h, the reaction was quenched by adding methanol (about2 mL) at 0° C. and the resulting mixture was stirred at room temperaturefor 15 min and then concentrated in vacuo. Dichloromethane (30 mL) andwater (30 mL) were added to the residue and this mixture was mixedthoroughly. The layers were separated and the aqueous layer wasextracted with dichloromethane (20 mL). The organic layers werecombined, dried (Na₂SO₄), filtered and the solvent was removed in vacuoto give the title compound as a brown foamy solid.

Preparation 24 Methyl4-(3-{[3-(Biphenyl-2-ylcarbamoyloxy)-3-methylbutyl]methylamino}propionylamino)-5-chloro-2-methoxybenzoate

To the crude product from Preparation 23 is added the debenzylatedproduct of Preparation 9 (4.5 mmol) and a mixture of THF (22.5 mL) andmethanol (2.5 mL). This mixture is heated at 50° C. with stirring for 16h and then the solvent is removed in vacuo. The residue ischromatographed to give the title compound.

Preparation 25 Biphenyl-2-ylcarbamic Acid3-{[2-(2-Chloro-4-hydroxymethyl-5-methoxy-phenylcarbamoyl)ethyl]methylamino}-1,1-dimethylpropylEster

To a solution of the product of Preparation 24 (1.45 mmol) in a mixtureof THF (4.5 mL) and methanol (0.5 mL) at 0° C. is added lithiumborohydride (32 mg, 1.45 mmol). The reaction mixture is allowed to warmto room temperature and is stirred for 41 h. The reaction is thenquenched by adding 1N aqueous hydrochloric acid at 0° C. until no morebubbling is observed and this mixture is stirred for 10 min. The solventis removed in vacuo and the residue is dissolved in acetonitrile (about2 mL). This solution is purified by prep-RP-HPLC. The appropriatefractions are collected and combined and lyophilized to give the titlecompound as a trifluoroacetate salt. This salt is treated with isopropylacetate (10 mL) and 1N aqueous sodium hydroxide (10 mL) and the organiclayer is collected, dried (Na₂SO₄), filtered and the solvent is removedin vacuo to give the title compound.

EXAMPLE 5 Biphenyl-2-ylcarbamic Acid3-{[2-(2-Chloro-4-{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-5-methoxyphenylcarbamoyl)ethyl]-methylamino}-1,1-dimethylpropylEster

Using the product of Preparation 25 and the procedures of Example 1, thetitle compound is prepared.

Preparation 26N-{2-Benzyloxy-5-[(R)-2-bromo-1-(tert-butyldimethylsilanyloxy)ethyl]phenyl}-formamide

(R)-2-Bromo-1-(3-formamido-4-benzyloxyphenyl)ethanol (9.9 g, 28 mmol)was dissolved in dimethylformamide (36 mL). Imidazole (2.3 g, 34 mmol)and tert-butyldimethylsilyl chloride (4.7 g, 31 mmol) were added. Thesolution was stirred under nitrogen atmosphere for 72 h. Additionalimidazole (0.39 g, 5.7 mmol) and tert-butyldimethylsilyl chloride (0.64g, 4.3 mmol) were added and the reaction was stirred for an additional20 h. The reaction mixture was then diluted with a mixture of isopropylacetate (53 mL) and hexanes (27 mL) and transferred to a separatoryfunnel. The organic layer was washed twice with a mixture of water (27mL) and saturated aqueous sodium chloride (27 mL) followed by a finalwash with saturated aqueous sodium chloride (27 mL). The organic layerwas dried over sodium sulfate. Silica gel (23.6 g) and hexanes (27 mL)were added and the suspension was stirred for 10 min. The solids wereremoved by filtration and the filtrate concentrated under vacuum. Theresidue was crystallized from hexanes (45 mL) to afford 8.85 g (19 mmol,68%) of the title compound as a solid. MS m/z: [M+H⁺] calcd forC₂₂H₃₀NO₃SiBr 464.1; found 464.2.

The starting material,(R)-2-bromo-1-(3-formamido-4-benzyloxyphenyl)ethanol, can be prepared asdescribed in U.S. Pat. No. 6,268,533 B1; or R. Hett et al., OrganicProcess Research and Development, 1998, 2:96-99; or using proceduressimilar to those described in Hong et al., Tetrahedron Lett., 1994,35:6631; or similar to those described in U.S. Pat. No. 5,495,054.

By using this compound in the examples above, additional compounds ofthis invention can be prepared.

Preparation 276-(2-Bromo-(R)-1-tert-butyldimethylsilyloxy)ethyl-2,2-dimethyl-1,3-benzodioxan(a) 6-Bromo-2,2-dimethyl-4H-benzo[1,3]dioxine

To 5-bromo-2-hydroxybenzyl alcohol (93 g, 0.46 mol, available fromSigma-Aldrich) in 2.0 L of 2,2-dimethoxypropane was added 700 mL ofacetone, followed by zinc chloride (170 g). After stirring for 18 hours,1.0 M aqueous sodium hydroxide was added until the aqueous phase wasbasic. Diethyl ether (1.5 L) was added to the slurry and the organicphase was decanted into a separatory funnel. The organic phase waswashed with brine, dried over Na₂SO₄, filtered and concentrated underreduced pressure to give the title compound as an oil.

(b) 6Acetyl-2,2-dimethyl-4H-benzo[1,3]dioxine

To the product of step (a) (110 g, 0.46 mol) in 1.0 L of THF at −78° C.was added 236 mL (0.51 mol) of 2.14 M n-butyllithium in hexanes via adropping funnel. After 30 minutes, N-methyl-N-methoxy acetamide (71 g,0.69 mol, available from TCI) was added. After 2 hours, the reactionmixture was quenched with water, diluted with 2.0 L of 1.0 M aqueousphosphate buffer (pH=7.0) and extracted once with diethyl ether. Thediethyl ether phase was washed once with brine, dried over Na₂SO₄,filtered and concentrated under reduced pressure to give a light orangeoil. The oil was dissolved in a minimum volume of ethyl acetate, dilutedwith hexanes, and to give the title compound as a crystalline solid.

(c) 6-Bromoacetyl-2,2-dimethyl-4H-benzo[1,3]dioxine

To the product of step (b) (23.4 g, 0.113 mol) in 600 mL of THF at −78°C. was added 135 mL of 1.0 M sodium hexamethyldisilazane in THF(Sigma-Aldrich). After 1 hour, trimethylsilyl chloride (15.8 mL, 0.124mol) was added. After another 30 minutes, bromine (5.82 mL, 0.113 mol)was added. After 10 minutes, the reaction was quenched by diluting thereaction mixture with diethyl ether and pouring it onto 500 mL of 5%aqueous Na₂SO₃ premixed with 500 mL of 5% aqueous NaHCO₃. The phaseswere separated and the organic phase was washed with brine, dried overNa₂SO₄, filtered and concentrated under reduced pressure to give thetitle compound as an oil that solidified upon storage in the freezer.

(d) (R)-2-Bromo-1-(2,2-dimethyl-4H-benzo[1,3]dioxin-6-yl)ethanol

To the product of step (c) (10 g, 35.1 mmol) in 100 mL of THF was addedthe solid catalyst of Preparation 13, step (c)(1) (0.97 g, 3.5 mmol).The solution was cooled to between −20° C. and −10° C. and BH₃-THF (35mL, 35 mmol) diluted with 50 mL THF was added dropwise via a droppingfunnel. After the addition was complete, the reaction mixture wasallowed to warm to ambient temperature. After 30 minutes, the reactionmixture was quenched by slow addition of 50 mL of methanol and thenconcentrated to a thick oil. The oil was purified by silica gelchromatography eluted with 1:2 ethyl acetate/hexanes. The fractions werecombined and concentrated to give the title compound as an off-whitesolid.

(e)[(R)-2-Bromo-1-(2,2-dimethyl-4H-benzo[1,3]dioxin-6yl)ethoxy]-tert-butyldimethylsilane

To the product of step (d) (10 g, 34.8 mmol) and imidazole (4.7 g, 69.7mmol) dissolved in 100 mL DMF was added tert-butyldimethylsilyl chloride(5.78 g, 38.3 mmol). The reaction mixture was stirred for 18 hours. Thereaction mixture was then partitioned between 200 mL of saturated sodiumchloride and 200 mL of diethyl ether. The aqueous layer was extractedwith 200 mL of diethyl ether. The organic layers were then combined,washed with saturated sodium chloride (3×100 mL), dried over MgSO₄ andconcentrated. The product was purified by silica gel chromatography,eluting with hexanes followed by 5% ethyl acetate in hexanes. Thedesired fractions were combined and concentrated to give the titlecompound as an oil.

By using this compound in the examples above, additional compounds ofthis invention can be prepared.

Preparation A Cell Culture and Membrane Preparation From CellsExpressing Human β₁, β₂ or β³ Adrenergic Receptors

Chinese hamster ovarian (CHO) cell lines stably expressing cloned humanβ₁, β₂ or β₃ adrenergic receptors, respectively, were grown to nearconfluency in Hams F-12 media with 10% FBS in the presence of 500 μg/mLGeneticin. The cell monolayer was lifted with 2 mM EDTA in PBS. Cellswere pelleted by centrifugation at 1,000 rpm, and cell pellets wereeither stored frozen at −80° C. or membranes were prepared immediatelyfor use. For preparation of β₁ and β₂ receptor expressing membranes,cell pellets were re-suspended in lysis buffer (10 mM HEPES/HCl, 10 mMEDTA, pH 7.4 at 4° C.) and homogenized using a tight-fitting Dounceglass homogenizer (30 strokes) on ice. For the more protease-sensitiveβ₃ receptor expressing membranes, cell pellets were homogenated in lysisbuffer (10 mM Tris/HCl, pH 7.4) supplemented with one tablet of“Complete Protease Inhibitor Cocktail Tablets with 2 mM EDTA” per 50 mLbuffer (Roche Catalog No. 1697498, Roche Molecular Biochemicals,Indianapolis, Ind.). The homogenate was centrifuged at 20,000× g, andthe resulting pellet was washed once with lysis buffer by re-suspensionand centrifugation as above. The final pellet was then re-suspended inice-cold binding assay buffer (75 mM Tris/HCl pH 7.4, 12.5 mM MgCl₂, 1mM EDTA). The protein concentration of the membrane suspension wasdetermined by the methods described in Lowry et al., 1951, Journal ofBiological Chemistry, 193, 265; and Bradford, Analytical Biochemistry,1976, 72, 248-54. All membranes were stored frozen in aliquots at −80°C. or used immediately.

Preparation B Cell Culture and Membrane Preparation From CellsExpressing Human M₁, M₂, M₃ and M₄ Muscarinic Receptors

CHO cell lines stably expressing cloned human hM₁, hM₂, hM₃ and hM₄muscarinic receptor subtypes, respectively, were grown to nearconfluency in HAM's F-12 media supplemented with 10% FBS and 250 μg/mLGeneticin. The cells were grown in a 5% CO₂, 37° C. incubator and liftedwith 2 mM EDTA in dPBS. Cells were collected by 5 minute centrifugationat 650× g, and cell pellets were either stored frozen at −80° C. ormembranes were prepared immediately for use. For membrane preparation,cell pellets were resuspended in lysis buffer and homogenized with aPolytron PT-2100 tissue disrupter (Kinematica AG; 20 seconds×2 bursts).Crude membranes were centrifuged at 40,000× g for 15 minutes at 4° C.The membrane pellet was then resuspended with re-suspension buffer andhomogenized again with the Polytron tissue disrupter. The proteinconcentration of the membrane suspension was determined by the methoddescribed in Lowry et al., 1951, Journal of Biochemistry, 193, 265. Allmembranes were stored frozen in aliquots at −80° C. or used immediately.Aliquots of prepared hM₅ receptor membranes were purchased directly fromPerkin Elmer and stored at −80° C. until use.

Assay Test Procedure A

Radioligand Binding Assay for Human β₁, β₂ and β₃ Adrenergic Receptors

Binding assays were performed in 96-well microtiter plates in a totalassay volume of 100 μL with 10-15 μg of membrane protein containing thehuman β₁, β₂ or β₃ adrenergic receptors in assay buffer (75 mM Tris/HClpH 7.4 at 25° C., 12.5 mM MgCl₂, 1 mM EDTA, 0.2% BSA). Saturationbinding studies for determination of K_(d) values of the radioligandwere done using [³H]-dihydroalprenolol (NET-720, 100 Ci/mmol,PerkinElmer Life Sciences Inc., Boston, Mass.) for the β₁ and β₂receptors and [¹²⁵I]-(−)-iodocyanopindolol (NEX-189, 220 Ci/mmol,PerkinElmer Life Sciences Inc., Boston, Mass.) at 10 or 11 differentconcentrations ranging from 0.01 nM to 20 nM. Displacement assays fordetermination of K_(i) values of test compounds were done with[³H]-dihydroalprenolol at 1 nM and [¹²⁵I]-(−)-iodocyanopindolol at 0.5nM for 10 or 11 different concentrations of test compound ranging from10 pM to 10 μM. Non-specific binding was determined in the presence of10 μM propranolol. Assays were incubated for 1 hour at 37° C., and thenbinding reactions were terminated by rapid filtration over GF/B for theβ₁ and β₂ receptors or GF/C glass fiber filter plates for the β₃receptors (Packard BioScience Co., Meriden, Conn.) presoaked in 0.3%polyethyleneimine. Filter plates were washed three times with filtrationbuffer (75 mM Tris/HCl pH 7.4 at 4° C., 12.5 mM MgCl₂, 1 mM EDTA) toremove unbound radioactivity. The plates were then dried and 50 μL ofMicroscint-20 liquid scintillation fluid (Packard BioScience Co.,Meriden, Conn.) was added and plates were counted in a Packard Topcountliquid scintillation counter (Packard BioScience Co., Meriden, Conn.).Binding data were analyzed by nonlinear regression analysis with theGraphPad Prism Software package (GraphPad Software, Inc., San Diego,Calif.) using the 3-parameter model for one-site competition. The curveminimum was fixed to the value for nonspecific binding, as determined inthe presence of 10 μM propranolol. K_(i) values for test compounds werecalculated from observed IC₅₀ values and the K_(d) value of theradioligand using the Cheng-Prusoff equation (Cheng Y, and Prusoff W H.,Biochemical Pharmacology, 1973, 22, 23, 3099-108).

In this assay, a lower K_(i) value indicates that a test compound has ahigher binding affinity for the receptor tested. Exemplified compound ofthis invention that were tested in this assay typically were found tohave a K_(i) value of less than about 300 nM for the β₂ adrenergicreceptor. For example, the compound of Example 1 was found to have K_(i)values of less than 300 nM.

If desired, the receptor subtype selectivity for a test compound can becalculated as the ratio of K_(i)(β₁)/K_(i)(β₂) or K_(i)(β₃)/K_(i)(β₂).Typically, compounds of this invention demonstrated greater binding atthe β₂ adrenergic receptor compared to the β₁ or β₃ adrenergic receptor,i.e. K_(i)(β₁) or K_(i)(β₃) is typically greater than K_(i)(β₂).Generally, compounds having selectivity for the β₂ adrenergic receptorover the β₁ or β₃ adrenergic receptors are preferred; especiallycompounds having a selectivity greater than about 5.

Assay Test Procedure B

Radioligand Binding Assay for Muscarinic Receptors

Radioligand binding assays for cloned human muscarinic receptors wereperformed in 96-well microtiter plates in a total assay volume of 100μL. CHO cell membranes stably expressing either the hM₁, hM₂, hM₃, hM₄or hM₅ muscarinic subtype were diluted in assay buffer to the followingspecific target protein concentrations (μg/well): 10 μg for hM₁, 10-15μg for hM₂, 10-20 μg for hM₃, 10-20 μg for hM₄, and 10-12 μg for hM₅ toget similar signals (cpm). The membranes were briefly homogenized usinga Polytron tissue disruptor (10 seconds) prior to assay plate addition.Saturation binding studies for determining K_(D) values of theradioligand were performed using L-[N-methyl-³H]scopolamine methylchloride ([³H]—NMS) (TRK666, 84.0 Ci/mmol, Amersham Pharmacia Biotech,Buckinghamshire, England) at concentrations ranging from 0.001 nM to 20nM. Displacement assays for determination of K_(i) values of testcompounds were performed with [³H]—NMS at 1 nM and eleven different testcompound concentrations. The test compounds were initially dissolved toa concentration of 400 μM in dilution buffer and then serially diluted5× with dilution buffer to final concentrations ranging from 10 μM to100 μM. The addition order and volumes to the assay plates were asfollows: 25 μL radioligand, 25 μL diluted test compound, and 50 μLmembranes. Assay plates were incubated for 60 minutes at 37° C. Bindingreactions were terminated by rapid filtration over GF/B glass fiberfilter plates (PerkinElmer Inc., Wellesley, Mass.) pre-treated in 1%BSA. Filter plates were rinsed three times with wash buffer (10 mMHEPES) to remove unbound radioactivity. The plates were then air driedand 50 μL Microscint-20 liquid scintillation fluid (PerkinElmer Inc.,Wellesley, Mass.) was added to each well. The plates were then countedin a PerkinElmer Topcount liquid scintillation counter (PerkinElmerInc., Wellesley, Mass.). Binding data were analyzed by nonlinearregression analysis with the GraphPad Prism Software package (GraphPadSoftware, Inc., San Diego, Calif.) using the one-site competition model.K_(i) values for test compounds were calculated from observed IC₅₀values and the K_(D) value of the radioligand using the Cheng-Prusoffequation (Cheng Y; Prusoff W H. (1973) Biochemical Pharmacology,22(23):3099-108). K_(i) values were converted to pK_(i) values todetermine the geometric mean and 95% confidence intervals. These summarystatistics were then converted back to K_(i) values for data reporting.

In this assay, a lower K_(i) value indicates that the test compound hasa higher binding affinity for the receptor tested. Exemplified compoundof this invention that were tested in this assay typically were found tohave a K_(i) value of less than about 300 nM for the M₃ muscarinicreceptor. For example, the compound of Example 1 was found to have K_(i)values of less than 300 nM for the M₃ muscarinic receptor.

Assay Test Procedure C

Whole-cell cAMP Flashplate Assay in CHO Cell Lines HeterologouslyExpressing Human β₁, β₂ or β₃ Adrenergic Receptors

cAMP assays are performed in a radioimmunoassay format using theFlashplate Adenylyl Cyclase Activation Assay System with [¹²⁵I]-cAMP(NEN SMP004, PerkinElmer Life Sciences Inc., Boston, Mass.), accordingto the manufacturers instructions. For the determination of P receptoragonist potency (EC₅₀), CHO-K1 cell lines stably expressing cloned humanβ₁, β₂ or β₃ adrenergic receptors are grown to near confluency in HAM'sF-12 media supplemented with 10% FBS and Geneticin (250 μg/mL). Cellsare rinsed with PBS and detached in dPBS (Dulbecco's Phosphate BufferedSaline, without CaCl₂ and MgCl₂) containing 2 mM EDTA or Trypsin-EDTAsolution (0.05% trypsin/0.53 mM EDTA). After counting cells in Coultercell counter, cells are pelleted by centrifugation at 1,000 rpm andre-suspended in stimulation buffer containing IBMX (PerkinElmer Kit)pre-warmed to room temperature to a concentration of 1.6×10⁶ to 2.8×10⁶cells/mL. About 60,000 to 80,000 cells per well are used in this assay.Test compounds (10 mM in DMSO) are diluted into PBS containing 0.1% BSAin Beckman Biomek-2000 and tested at 11 different concentrations rangingfrom 100 μM to 1 μM. Reactions are incubated for 10 min at 37° C. andstopped by adding 100 μL of cold detection buffer containing [¹²⁵I]-cAMP(NEN SMP004, PerkinElmer Life Sciences, Boston, Mass.). The amount ofcAMP produced (pmol/well) is calculated based on the counts observed forthe samples and cAMP standards as described in the manufacturer's usermanual. Data are analyzed by nonlinear regression analysis with theGraphPad Prism Software package (GraphPad Software, Inc., San Diego,Calif.) with the sigmoidal equation. The Cheng-Prusoff equation (ChengY, and Prusoff W H., Biochemical Pharmacology, 1973, 22, 23, 3099-108)is used to calculate the EC50 values.

In this assay, a lower EC₅₀ value indicates that the test compound has ahigher functional activity at the receptor tested. When tested in thisassay, compound of this invention are expected to have a EC₅₀ value ofless than about 300 nM for the β₂ adrenergic receptor.

If desired, the receptor subtype selectivity for a test compound can becalculated as the ratio of EC₅₀(β₁)/EC₅₀(β₂) or EC₅₀(β₃)/EC₅₀(β₂).Typically, compounds of this invention demonstrated greater functionalactivity at the β₂ adrenergic receptor compared to the β₁ β₃ adrenergicreceptor, i.e. EC₅₀(β₁) or EC₅₀(β₃) is typically greater than EC₅₀(β₂).Generally, compounds having selectivity for the β₂ adrenergic receptorover the β₁ or β₃ adrenergic receptors are preferred; especiallycompounds having a selectivity greater than about 5; and in particular,greater than about 10.

Assay Test Procedure D

Functional Assays of Antagonism for Muscarinic Receptor Subtypes

A. Blockade of Agonist-Mediated Inhibition of cAMP Accumulation

In this assay, the functional potency of a test compound is determinedby measuring the ability of the test compound to blockoxotremorine-inhibition of forskolin-mediated cAMP accumulation inCHO-K1 cells expressing the hM₂ receptor. cAMP assays are performed in aradioimmunoassay format using the Flashplate Adenylyl Cyclase ActivationAssay System with ¹²⁵I-cAMP (NEN SMP004B, PerkinElmer Life SciencesInc., Boston, Mass.), according to the manufacturer's instructions.Cells are rinsed once with dPBS and lifted with Trypsin-EDTA solution(0.05% trypsin/0.53 mM EDTA) as described in the Cell Culture andMembrane Preparation section above. The detached cells are washed twiceby centrifugation at 650× g for five minutes in 50 mL dPBS. The cellpellet is then re-suspended in 10 mL dPBS, and the cells are countedwith a Coulter Z1 Dual Particle Counter (Beckman Coulter, Fullerton,Calif.). The cells are centrifuged again at 650× g for five minutes andre-suspended in stimulation buffer to an assay concentration of1.6×10⁶-2.8×10⁶ cells/mL.

The test compound is initially dissolved to a concentration of 400 μM indilution buffer (dPBS supplemented with 1 mg/mL BSA (0.1%)), and thenserially diluted with dilution buffer to final molar concentrationsranging from 100 μM to 0.1 nM. Oxotremorine is diluted in a similarmanner.

To measure oxotremorine inhibition of adenylyl cyclase (AC) activity, 25μL forskolin (25 μM final concentration diluted in dPBS), 25 μL dilutedoxotremorine, and 50 μL cells are added to agonist assay wells. Tomeasure the ability of a test compound to block oxotremorine-inhibitedAC activity, 25 μL forskolin and oxotremorine (25 μM and 5 μM finalconcentrations, respectively, diluted in dPBS), 25 μL diluted testcompound, and 50 μL cells are added to remaining assay wells.

Reactions are incubated for 10 minutes at 37° C. and stopped by additionof 100 μL ice-cold detection buffer. Plates are sealed, incubatedovernight at room temperature and counted the next morning on aPerkinElmer TopCount liquid scintillation counter (PerkinElmer Inc.,Wellesley, Mass.). The amount of cAMP produced (pmol/well) is calculatedbased on the counts observed for the samples and cAMP standards, asdescribed in the manufacturer's user manual. Data is analyzed bynonlinear regression analysis with the GraphPad Prism Software package(GraphPad Software, Inc., San Diego, Calif.) using the non-linearregression, one-site competition equation. The Cheng-Prusoff equation isused to calculate the K_(i), using the EC₅₀ of the oxotremorineconcentration-response curve and the oxotremorine assay concentration asthe K_(D) and [L], respectively.

In this assay, a lower K_(i) value indicates that the test compound hasa higher functional activity at the receptor tested. When tested in thisassay, compounds of this invention are expected to have a K_(i) value ofless than about 300 nM for blockade of oxotremorine-inhibition offorskolin-mediated cAMP accumulation in CHO-K1 cells expressing the hM₂receptor.

B. Blockade of Agonist-Mediated [³⁵S]GTPγS Binding

In a second functional assay, the functional potency of test compoundscan be determined by measuring the ability of the compounds to blockoxotremorine-stimulated [³⁵S]GTPγS_binding in CHO-K1 cells expressingthe hM₂ receptor.

At the time of use, frozen membranes are thawed and then diluted inassay buffer with a final target tissue concentration of 5-10 μg proteinper well. The membranes are briefly homogenized using a Polytron PT-2100tissue disrupter and then added to the assay plates.

The EC₉₀ value (effective concentration for 90% maximal response) forstimulation of [³⁵S]GTPγS binding by the agonist oxotremorine isdetermined in each experiment.

To determine the ability of a test compound to inhibitoxotremorine-stimulated [³⁵S]GTPγS binding, the following is added toeach well of 96 well plates: 25 μL of assay buffer with [³⁵S]GTPγS (0.4nM), 25 μL of oxotremorine(EC₉₀) and GDP (3 uM), 25 μL of diluted testcompound and 25 μL CHO cell membranes expressing the hM₂ receptor. Theassay plates are then incubated at 37° C. for 60 minutes. The assayplates are filtered over 1% BSA-pretreated GF/B filters using aPerkinElmer 96-well harvester. The plates are rinsed with ice-cold washbuffer for 3×3 seconds and then air or vacuum dried. Microscint-20scintillation liquid (50 μL) is added to each well, and each plate issealed and radioactivity counted on a Topcounter (PerkinElmer). Data areanalyzed by nonlinear regression analysis with the GraphPad PrismSoftware package (GraphPad Software, Inc., San Diego, Calif.) using thenon-linear regression, one-site competition equation. The Cheng-Prusoffequation is used to calculate the K_(i), using the IC₅₀ values of theconcentration-response curve for the test compound and the oxotremorineconcentration in the assay as the K_(D) and [L], ligand concentration,respectively.

In this assay, a lower K_(i) value indicates that the test compound hasa higher functional activity at the receptor tested. When tested in thisassay, compounds of this invention are expected to have a K_(i) value ofless than about 300 nM for blockade of oxotremorine-stimulated[³⁵S]GTPγS_binding in CHO-K1 cells expressing the hM₂ receptor.

C. Blockade of Agonist-Mediated Calcium Release via FLIPR Assays

Muscarinic receptor subtypes (M₁, M₃ and M₅ receptors), which couple toG_(q) proteins, activate the phospholipase C (PLC) pathway upon agonistbinding to the receptor. As a result, activated PLC hydrolyzesphosphatyl inositol diphosphate (PIP₂) to diacylglycerol (DAG) andphosphatidyl-1,4,5-triphosphate (IP₃), which in turn generates calciumrelease from intracellular stores, i.e., endoplasmic and sarcoplasmicreticulum. The FLIPR (Molecular Devices, Sunnyvale, Calif.) assaycapitalizes on this increase in intracellular calcium by using a calciumsensitive dye (Fluo-4AM, Molecular Probes, Eugene, Oreg.) thatfluoresces when free calcium binds. This fluorescence event is measuredin real time by the FLIPR, which detects the change in fluorescence froma monolayer of cells cloned with human M₁ and M₃, and chimpanzee M₅receptors. Antagonist potency can be determined by the ability ofantagonists to inhibit agonist-mediated increases in intracellularcalcium.

For FLIPR calcium stimulation assays, CHO cells stably expressing thehM₁, hM₃ and cM₅ receptors are seeded into 96-well FLIPR plates thenight before the assay is done. Seeded cells are washed twice byCellwash (MTX Labsystems, Inc.) with FLIPR buffer (10 mM HEPES, pH 7.4,2 mM calcium chloride, 2.5 mM probenecid in Hank's Buffered SaltSolution (HBSS) without calcium and magnesium) to remove growth mediaand leaving 50 μL/well of FLIPR buffer. The cells are then incubatedwith 50 μL/well of 4 μM FLUO-4AM (a 2× solution was made) for 40 minutesat 37° C., 5% carbon dioxide. Following the dye incubation period, cellsare washed two times with FLIPR buffer, leaving a final volume of 50μL/well.

To determine antagonist potency, the dose-dependent stimulation ofintracellular Ca²⁺ release for oxotremorine is first determined so thatantagonist potency can later be measured against oxotremorinestimulation at an EC₉₀ concentration. Cells are first incubated withcompound dilution buffer for 20 minutes, followed by agonist addition,which is performed by the FLIPR. An EC₉₀ value for oxotremorine isgenerated according to the method detailed in the FLIPR measurement anddata reduction section below, in conjunction with the formulaEC_(F)=((F/100-F)^1/H)*EC₅₀. An oxotremorine concentration of 3×EC_(F)is prepared in stimulation plates such that an EC₉₀ concentration ofoxotremorine is added to each well in the antagonist inhibition assayplates.

The parameters used for the FLIPR are: exposure length of 0.4 seconds,laser strength of 0.5 watts, excitation wavelength of 488 nm, andemission wavelength of 550 nm. Baseline is determined by measuring thechange in fluorescence for 10 seconds prior to addition of agonist.Following agonist stimulation, the FLIPR continuously measured thechange of fluorescence every 0.5 to 1 second for 1.5 minutes to capturethe maximum fluorescence change.

The change of fluorescence is expressed as maximum fluorescence minusbaseline fluorescence for each well. The raw data is analysed againstthe logarithm of drug concentration by nonlinear regression withGraphPad Prism (GraphPad Software, Inc., San Diego, Calif.) using thebuilt-in model for sigmoidal dose-response. Antagonist K_(i) values aredetermined by Prism using the oxotremorine EC₅₀ value as the K_(D) andthe oxotremorine EC₉₀ for the ligand concentration according to theCheng-Prusoff equation (Cheng & Prusoff, 1973).

In this assay, a lower K_(i) value indicates that the test compound hasa higher functional activity at the receptor tested. When tested in thisassay, compounds of this invention are expected to have a K_(i) value ofless than about 300 nM for blockade of agonist-mediated calcium releasein CHO cells stably expressing the hM₁, hM₃ and cM₅ receptors.

Assay Test Procedure E

Whole-cell cAMP Flashplate Assay With a Lung Epithelial Cell LineEndogenously Expressing Human β₂ Adrenergic Receptor

For the determination of agonist potencies and efficacies (intrinsicactivities) in a cell line expressing endogenous levels of the β₂adrenergic receptor, a human lung epithelial cell line (BEAS-2B) is used(ATCC CRL-9609, American Type Culture Collection, Manassas, Va.)(January B, et al., British Journal of Pharmacology, 1998, 123, 4,701-11). Cells are grown to 75-90% confluency in complete, serum-freemedium (LHC-9 MEDIUM containing Epinephrine and Retinoic Acid, cat #181-500, Biosource International, Camarillo, Calif.). The day before theassay, medium is switched to LHC-8 (no epinephrine or retinoic acid, cat# 141-500, Biosource International, Camarillo, Calif.). cAMP assays areperformed in a radioimmunoassay format using the Flashplate AdenylylCyclase Activation Assay System with [¹²⁵I]-cAMP (NEN SMP004,PerkinElmer Life Sciences Inc., Boston, Mass.), according to themanufacturers instructions.

On the day of the assay, cells are rinsed with PBS, lifted by scrapingwith 5 mM EDTA in PBS, and counted. Cells are pelleted by centrifugationat 1,000 rpm and re-suspended in stimulation buffer pre-warmed to 37° C.at a final concentration of 600,000 cells/mL. Cells are used at a finalconcentration of 100,000 to 120,000 cells/well in this assay. Testcompounds are serially diluted into assay buffer (75 mM Tris/HCl pH 7.4at 25° C., 12.5 mM MgCl₂, 1 mM EDTA, 0.2% BSA) in Beckman Biomek-2000.Test compounds are tested in the assay at 11 different concentrations,ranging from 10 μM to 10 pM. Reactions are incubated for 10 min at 37°C. and stopped by addition of 100 μL of ice-cold detection buffer.Plates are sealed, incubated over night at 4° C. and counted the nextmorning in a Topcount scintillation counter (Packard BioScience Co.,Meriden, Conn.). The amount of cAMP produced per mL of reaction iscalculated based on the counts observed for samples and cAMP standards,as described in the manufacturer's user manual. Data are analyzed bynonlinear regression analysis with the GraphPad Prism Software package(GraphPad Software, Inc., San Diego, Calif.) using the 4-parameter modelfor sigmoidal dose-response.

In this assay, a lower EC₅₀ value indicates that the test compound has ahigher functional activity at the receptor tested. When tested in thisassay, compounds of this invention are expected to have an EC₅₀ value ofless than about 300 nM for the β₂ adrenergic receptor.

If desired, test compound efficacy (% Eff) is calculated from the ratioof the observed Emax (TOP of the fitted curve) and the maximal responseobtained for isoproterenol dose response curve and is expressed as % Effrelative to isoproterenol. Compounds of this invention are expected todemonstrate a % Eff greater than about 40.

Assay Test Procedure F

Duration of Bronchoprotection In Guinea Pig Models ofAcetylcholine-Induced or Histamine-Induced Bronchoconstriction

These in vivo assays are used to assess the bronchoprotective effects oftest compounds exhibiting both muscarinic receptor antagonist and β₂adrenergic receptor agonist activity. To isolate muscarinic antagonistactivity in the acetylcholine-induced bronchoconstriction model, theanimals are administered propanolol, a compound that blocks β receptoractivity, prior to the administration of acetylcholine. Duration ofbronchoprotection in the histamine-induced bronchoconstriction modelreflects β₂ adrenergic receptor agonist activity.

Groups of 6 male guinea pigs (Duncan-Hartley (HsdPoc:DH) Harlan,Madison, Wis.) weighing between 250 and 350 g are individuallyidentified by cage cards. Throughout the study, animals are allowedaccess to food and water ad libitum.

Test compounds are administered via inhalation over 10 minutes in awhole-body exposure dosing chamber (R&S Molds, San Carlos, Calif.). Thedosing chambers are arranged so that an aerosol is simultaneouslydelivered to 6 individual chambers from a central manifold. Guinea pigsare exposed to an aerosol of a test compound or vehicle (WFI). Theseaerosols are generated from aqueous solutions using an LC Star NebulizerSet (Model 22F51, PARI Respiratory Equipment, Inc. Midlothian, Va.)driven by a mixture of gases (CO₂=5%, O₂=21% and N₂=74%) at a pressureof 22 psi. The gas flow through the nebulizer at this operating pressureis approximately 3 L/minute. The generated aerosols are driven into thechambers by positive pressure. No dilution air is used during thedelivery of aerosolized solutions. During the 10 minute nebulization,approximately 1.8 mL of solution is nebulized. This value is measuredgravimetrically by comparing pre- and post-nebulization weights of thefilled nebulizer.

The bronchoprotective effects of test compounds administered viainhalation are evaluated using whole body plethysmography at 1.5, 24, 48and 72 hours post-dose.

Forty-five minutes prior to the start of the pulmonary evaluation, eachguinea pig is anesthetized with an intramuscular injection of ketamine(43.75 mg/kg), xylazine (3.50 mg/kg) and acepromazine (1.05 mg/kg).After the surgical site is shaved and cleaned with 70% alcohol, a 2-3 cmmidline incision of the ventral aspect of the neck is made. Then, thejugular vein is isolated and cannulated with a saline-filledpolyethylene catheter (PE-50, Becton Dickinson, Sparks, Md.) to allowfor intravenous infusions of acetylcholine (Ach) or histamine in saline.The trachea is then dissected free and cannulated with a 14G teflon tube(#NE-014, Small Parts, Miami Lakes, Fla.). If required, anesthesia ismaintained by additional intramuscular injections of the aforementionedanesthetic mixture. The depth of anesthesia is monitored and adjusted ifthe animal responds to pinching of its paw or if the respiration rate isgreater than 100 breaths/minute.

Once the cannulations are completed, the animal is placed into aplethysmograph (#PLY3114, Buxco Electronics, Inc., Sharon, Conn.) and anesophageal pressure cannula (PE-160, Becton Dickinson, Sparks, Md.) isinserted to measure pulmonary driving pressure (pressure). The teflontracheal tube is attached to the opening of the plethysmograph to allowthe guinea pig to breathe room air from outside the chamber. The chamberis then sealed. A heating lamp is used to maintain body temperature andthe guinea pig's lungs are inflated 3 times with 4 mL of air using a 10mL calibration syringe (#5520 Series, Hans Rudolph, Kansas City, Mo.) toensure that the lower airways did not collapse and that the animal didnot suffer from hyperventilation.

Once it was determined that baseline values are within the range of0.3-0.9 mL/cm H₂O for compliance and within the range of 0.1-0.199 cmH₂O/mL per second for resistance, the pulmonary evaluation is initiated.A Buxco pulmonary measurement computer progam enabled the collection andderivation of pulmonary values.

Starting this program initiated the experimental protocol and datacollection. The changes in volume over time that occur within theplethysmograph with each breath are measured via a Buxco pressuretransducer. By integrating this signal over time, a measurement of flowis calculated for each breath. This signal, together with the pulmonarydriving pressure changes, which are collected using a Sensym pressuretransducer (#TRD4100), is connected via a Buxco (MAX 2270) preamplifierto a data collection interface (#'s SFT3400 and SFT3813). All otherpulmonary parameters are derived from these two inputs.

Baseline values are collected for 5 minutes, after which time the guineapigs are challenged with Ach or histamine. When evaluating themuscarinic antagonist effects, propanolol (5 mg/Kg, iv) (Sigma-Aldrich,St. Louis, Mo.) is administered 15 minutes prior to challenge with Ach.Ach (Sigma-Aldrich, St. Louis, Mo.) (0.1 mg/mL) is infused intravenouslyfor 1 minute from a syringe pump (sp210iw, World Precision Instruments,Inc., Sarasota, Fla.) at the following doses and prescribed times fromthe start of the experiment: 1.9 μg/minute at 5 minutes, 3.8 μg/minuteat 10 minutes, 7.5 μg/minute at 15 minutes, 15.0 μg/minute at 20minutes, 30 μg/minute at 25 minutes and 60 μg/minute at 30 minutes.Alternatively, bronchoprotection of test compounds is assessed in theacetylcholine challenge model without pretreatment with a beta blockingcompound.

When evaluating the β₂ adrenergic receptor agonist effects of testcompounds, histamine (25 μg/mL) (Sigma-Aldrich, St. Louis, Mo.) isinfused intravenously for 1 minute from a syringe pump at the followingdoses and prescribed times from the start of the experiment: 0.5μg/minute at 5 minutes, 0.9 μg/minute at 10 minutes, 1.9 μg/minute at 15minutes, 3.8 μg/minute at 20 minutes, 7.5 μg/minute at 25 minutes and 15μg/minute at 30 minutes. If resistance or compliance does not returnedto baseline values at 3 minutes following each Ach or histamine dose,the guinea pig's lungs are inflated 3 times with 4 mL of air from a 10mL calibration syringe. Recorded pulmonary parameters includerespiration frequency (breaths/minute), compliance (mL/cm H₂O) andpulmonary resistance (cm H₂O/mL per second). Once the pulmonary functionmeasurements are completed at minute 35 of this protocol, the guinea pigis removed from the plethysmograph and euthanized by carbon dioxideasphyxiation.

The data are evaluated in one of two ways:

(a) Pulmonary resistance (R_(L), cm H₂O/mL per second) is calculatedfrom the ratio of “change in pressure” to “the change in flow.” TheR_(L) response to ACh (60 μg/min, 1H) is computed for the vehicle andthe test compound groups. The mean ACh response in vehicle-treatedanimals, at each pre-treatment time, is calculated and used to compute %inhibition of ACh response, at the corresponding pre-treatment time, ateach test compound dose. Inhibition dose-response curves for ‘R_(L)’were fitted with a four parameter logistic equation using GraphPadPrism, version 3.00 for Windows (GraphPad Software, San Diego, Calif.)to estimate bronchoprotective ID₅₀ (dose required to inhibit the ACh (60μg/min) bronchocontrictor response by 50%). The equation used is asfollows:Y=Min+(Max−Min)/(1+10^(((log ID50−X)*Hillslope)))where X is the logarithm of dose, Y is the response (% Inhibition of AChinduced increase in R_(L)). Y starts at Min and approachesasymptotically to Max with a sigmoidal shape. (b) The quantity PD₂,which is defined as the amount of Ach or histamine needed to cause adoubling of the baseline pulmonary resistance, is calculated using thepulmonary resistance values derived from the flow and the pressure overa range of Ach or histamine challenges using the following equation(which was derived from an equation used to calculate PC₂₀ valuesdescribed in American Thoracic Society. Guidelines for methacholine andexercise challenge testing—1999. Am J Respir Crit Care Med. 2000; 161:309-329):

${PD}_{2} = {{antilog}\mspace{11mu}\left\lbrack {{\log\mspace{14mu} C_{1}} + \frac{\left( {{\log\mspace{14mu} C_{2}} - {\log\mspace{14mu} C_{1}}} \right)\left( {{2R_{0}} - R_{1}} \right)}{R_{2} - R_{1}}} \right\rbrack}$

where:

-   -   C₁=concentration of Ach or histamine preceding C₂    -   C₂=concentration of Ach or histamine resulting in at least a        2-fold increase in pulmonary resistance (R^(L))    -   R₀=Baseline R_(L) value    -   R₁=R_(L) value after C₁    -   R₂=R_(L) value after C₂

Statistical analysis of the data is performed using a twotailed—Students t-test. A P-value <0.05 was considered significant.

Exemplified compounds of this invention are expected to produce adose-dependent bronchoprotective effect against MCh-inducedbronchoconstriction and His-induced bronchoconstriction. Test compoundshaving a potency (ID₅₀ at 1.5 h post-dose) of less than about 300 μg/mLfor ACh-induced bronchoconstriction and less than about 300 μg/mL forHis-induced bronchoconstriction in this assay are generally preferred.Additionally, test compounds having a duration (PD T_(1/2)) ofbrochoprotective activity of at least about 24 hours in this assay aregenerally preferred.

Assay Test Procedure G

Einthoven Model for Measuring Changes in Ventilation in Guinea Pigs

The bronchodilator activity of test compounds is evaluated in ananesthetized guinea pig model (the Einthoven model), which usesventilation pressure as a surrogate measure of airway resistance. See,for example, Einthoven (1892) Pfugers Arch. 51: 367-445; and Mohammed etal. (2000) Pulm Pharmacol Ther. 13(6):287-92. In this model, muscarinicantagonist and β₂ agonist activity is assessed by determining theprotective effects against methacholine (MCh) and histamine(His)-induced bronchoconstriction.

This assay is conducted using Duncan-Hartley guinea pigs (Harlan,Indianapolis, Ind.), weighing between 300 and 400 g.

The test compound or vehicle (i.e., sterile water) is dosed byinhalation (1H) over a 10 minute time period in a whole body exposuredosing chamber (R+S Molds, San Carlos, Calif.) using 5 mL of dosingsolution. Animals are exposed to an aerosol, which is generated from anLC Star Nebulizer Set (Model 22F51, PARI Respiratory Equipment, Inc.Midlothian, Va.) driven by Bioblend a mixture of gasses (5% CO₂; 21% O₂;and 74% N₂) at a pressure of 22 psi. Pulmonary function is evaluated atvarious time-points after inhalation dosing.

Forty five minutes prior to the start of pulmonary function evaluation,the guinea pigs are anesthetized with an intramuscular (1M) injection ofa mixture of ketamine (13.7 mg/kg/xylazine (3.5 mg/kg)/acepromazine(1.05 mg/kg). A supplemental dose of this mixture (50% of initial dose)is administered as needed. The jugular vein and carotid artery areisolated and cannulated with saline-filled polyethylene catheters(micro-renathane and PE-50, respectively, Beckton Dickinson, Sparks,Md.). The carotid artery is connected to a pressure transducer to allowthe measurement of blood pressure and the jugular vein cannula is usedfor IV injection of either MCh or His. The trachea is then dissectedfree and cannulated with a 14G needle (#NE-014, Small Parts, MiamiLakes, Fla.). Once the cannulations are complete, the guinea pigs areventilated using a respirator (Model 683, Harvard Apparatus, Inc., MA)set at a stroke volume of 1 mL/100 g body weight but not exceeding 2.5mL volume, and at a rate of 100 strokes per minute. Ventilation pressure(VP) is measured in the tracheal cannula using a Biopac transducer thatis connected to a Biopac (TSD 137C) pre-amplifier. Body temperature ismaintained at 37° C. using a heating pad. Prior to initiating datacollection, pentobarbital (25mg/kg) is administered intraperitoneally(IP) to suppress spontaneous breathing and obtain a stable baseline. Thechanges in VP are recorded on a Biopac Windows data collectioninterface. Baseline values are collected for at least 5 minutes, afterwhich time guinea pigs are challenged IV non-cumulatively with 2-foldincremental doses of the bronchoconstrictor (MCh or His). When MCh isused as the bronchoconstrictor agent, animals are pre-treated withpropranolol (5 mg/kg, IV) to isolate the antimuscarinic effects of thetest compound. Changes in VP are recorded using the Acknowledge DataCollection Software (Santa Barbara, Calif.). After the completion ofstudy, the animals are euthanized.

Change in VP is measured in cm of water. Change in VP (cm H₂O)=peakpressure (after bronchoconstrictor challenge)−peak baseline pressure.The dose-response curve to MCh or His is fitted to a four parameterlogistic equation using GraphPad Prism, version 3.00 for Windows(GraphPad Software, San Diego, Calif.). The equation used is as follows:Y=Min+(Max−Min)/(1+10^(((log ID50−X)*Hillslope)))where X is the logarithm of dose, Y is the response. Y starts at Min andapproaches asymptotically to Max with a sigmoidal shape.

The percent inhibition of the bronchoconstrictor response to asubmaximal dose of MCh or His is calculated at each dose of the testcompound using the following equation: % Inhibition ofresponse=100−((peak pressure (after bronchoconstrictor challenge,treated)−peak baseline pressure (treated)*100%/(peak pressure (afterbronchoconstrictor challenge, water)−peak baseline pressure (water)).Inhibition curves are fitted using the four parameter logistic equationfrom GraphPad software. ID₅₀ (dose required to produce 50% inhibition ofthe bronchoconstrictor response) and Emax (maximal inhibition) are alsoestimated wherever appropriate.

The magnitude of bronchoprotection at different time-points afterinhalation of the test compound is used to estimate the pharmacodynamichalf-life (PD T_(1/2)). PD T_(1/2) is determined using a non-linearregression fit using a one-phase exponential decay equation (GraphPadPrism, Version 4.00): Y=Span*exp(−K*X)+Plateau; Starts at Span+Plateauand decays to Plateau with a rate constant K. The PD T_(1/2)=0.69/K.Plateau is constrained to 0.

Exemplified compounds of this invention are expected to produce adose-dependent bronchoprotective effect against MCh-inducedbronchoconstriction and His-induced bronchoconstriction. Generally, testcompounds having an ID₅₀ less than about 300 μg/mL for MCh-inducedbronchoconstriction and an ID₅₀ less than about 300 μg/mL forHis-induced bronchoconstriction at 1.5 hours post-dose in this assay arepreferred. Additionally, test compounds having a duration (PD T_(1/2))of brochoprotective activity of at least about 24 hours in this assayare generally preferred.

Assay Test Procedure H

Inhalation Guinea Pig Salivation Assay

Guinea pigs (Charles River, Wilmington, Mass.) weighing 200-350 g areacclimated to the in-house guinea pig colony for at least 3 daysfollowing arrival. Test compound or vehicle are dosed via inhalation(1H) over a 10 minute time period in a pie shaped dosing chamber (R+SMolds, San Carlos, Calif.). Test solutions are dissolved in sterilewater and delivered using a nebulizer filled with 5.0 mL of dosingsolution. Guinea pigs are restrained in the inhalation chamber for 30minutes. During this time, guinea pigs are restricted to an area ofapproximately 110 sq. cm. This space is adequate for the animals to turnfreely, reposition themselves, and allow for grooming. Following 20minutes of acclimation, guinea pigs are exposed to an aerosol generatedfrom a LS Star Nebulizer Set (Model 22F51, PARI Respiratory Equipment,Inc. Midlothian, Va.) driven by house air at a pressure of 22 psi. Uponcompletion of nebulization, guinea pigs are evaluated at 1.5, 6, 12, 24,48, or 72 hrs after treatment.

Guinea pigs are anesthetized one hour before testing with anintramuscular (1M) injection of a mixture of ketamine 43.75 mg/kg,xylazine 3.5 mg/kg, and acepromazine 1.05 mg/kg at an 0.88 mL/kg volume.Animals are placed ventral side up on a heated (37° C.) blanket at a 20degree incline with their head in a downward slope. A 4-ply 2×2 inchgauze pad (Nu-Gauze General-use sponges, Johnson and Johnson, Arlington,Tex.) is inserted in the guinea pig's mouth. Five minutes later, themuscarinic agonist pilocarpine (3.0 mg/kg, s.c.) is administered and thegauze pad is immediately discarded and replaced by a new pre-weighedgauze pad. Saliva is collected for 10 minutes, at which point the gauzepad is weighed and the difference in weight recorded to determine theamount of accumulated saliva (in mg). The mean amount of salivacollected for animals receiving the vehicle and each dose of testcompound is calculated. The vehicle group mean is considered to be 100%salivation. Results are calculated using result means (n=3 or greater).Confidence intervals (95%) are calculated for each dose at each timepoint using two-way ANOVA. This model is a modified version of theprocedure described in Rechter, “Estimation of anticholinergic drugeffects in mice by antagonism against pilocarpine-induced salivation”Ata Pharmacol Toxicol, 1996, 24:243-254.

The mean weight of saliva in vehicle-treated animals, at eachpre-treatment time, is calculated and used to compute % inhibition ofsalivation, at the corresponding pre-treatment time, at each dose. Theinhibition dose-response data are fitted to a four parameter logisticequation using GraphPad Prism, version 3.00 for Windows (GraphPadSoftware, San Diego, Calif.) to estimate anti-sialagogue ID₅₀(doserequired to inhibit 50% of pilocarpine-evoked salivation). The equationused is as follows:Y=Min+(Max−Min)/(1+10^(((log ID50−X)*Hillslope)))where X is the logarithm of dose, Y is the response (% inhibition ofsalivation). Y starts at Min and approaches asymptotically to Max with asigmoidal shape.

The ratio of the anti-sialagogue ID₅₀ to bronchoprotective ID₅₀ is usedto compute the apparent lung-selectivity index of the test compound.Generally, compounds having an apparent lung-selectivity index greaterthan about 5 are preferred.

While the present invention has been described with reference tospecific aspects or embodiments thereof, it will be understood by thoseof ordinary skilled in the art that various changes can be made orequivalents can be substituted without departing from the true spiritand scope of the invention. Additionally, to the extent permitted byapplicable patent statutes and regulations, all publications, patentsand patent applications cited herein are hereby incorporated byreference in their entirety to the same extent as if each document hadbeen individually incorporated by reference herein.

1. A compound of formula I:

wherein W represents O or NW^(a); where W^(a) is hydrogen or(1-4C)alkyl; R¹ is (6-10C)aryl, (2-9C)heteroaryl containing from 1 to 4heteroatoms independently selected from oxygen, nitrogen and sulfur or(3-7C)cycloalkyl; wherein the aryl, heteroaryl or cycloalkyl group isunsubstituted or substituted with from 1 to 3 substituents independentlyselected from (1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl,(3-6C)cycloalkyl, (6-10C)aryl, (2-9C)heteroaryl, (3-5C)heterocyclic,cyano, halo, —OR^(1a), —C(O)OR^(1b), —SR^(1c), —S(O)R^(1d),—S(O)₂R^(1e), —NR^(1f)R^(1g), —C(O)R^(1h), —NR^(1i)C(O)OR^(1j),—OC(O)NR^(1k)R^(1l), —NR^(1m)C(O)R^(1n), —C(O)NR^(1o)R^(1p) and—NHS(O)₂R^(1q) where each of R^(1a), R^(1b), R^(1c), R^(1d), R^(1e),R^(1f), R^(1g), R^(1h), R^(1i), R^(1j), R^(1k), R^(1l), R^(1m), R^(1n),R^(1o), R^(1p) and R^(1q) is independently selected from hydrogen,(1-4C)alkyl or phenyl-(1-4C)alkyl; and where each (3-6C)cycloalkyl,(6-10C)aryl, (2-9C)heteroaryl and (3-5C)heterocyclic group isunsubstituted or substituted with from 1 to 3 substituents selected from(1-4C)alkyl, halo and —OR^(1a); and where each alkyl group present in R¹is unsubstituted or substituted with from 1 to 3 fluoro substituents;each R² is independently selected from (1-4C)alkyl, (2-4C)alkenyl,(2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(2a), —C(O)OR^(2b),—SR^(2c), —S(O)R^(2d), S(O)₂R^(2e) and —NR^(2f)R^(2g); where each ofR^(2a), R^(2b), R^(2c), R^(2d), R^(2e), R^(2f) and R^(2g) isindependently selected from hydrogen, (1-4C)alkyl or phenyl-(1-4C)alkyl;and where each alkyl group present in R² is unsubstituted or substitutedwith from 1 to 3 fluoro substituents; R³ is selected from hydrogen,(1-6C)alkyl, (2-6C)alkenyl, (2-6C)alkynyl, or (3-6C)cycloalkyl, wherethe alkyl group is unsubstituted or substituted with from 1 to 3 fluorosubstituents; R⁴ represents a divalent hydrocarbon group containing from4 to 28 carbon atoms and optionally containing from 1 to 10 heteroatomsselected independently from halo, oxygen, nitrogen and sulfur, providedthat the number of contiguous atoms in the shortest chain between thetwo nitrogen atoms to which R⁴ is attached is in the range of from 4 to16; R⁵ represents hydrogen or (1-4C)alkyl; R⁶ and R⁷ together form—N(R^(7a))C(O)C(R^(7b))═C(R^(7c))—, —C(R^(7d))═C(R^(7e))C(O)N(R^(7f))—,—N(R^(7g))C(O)CR^(7h)R^(7i)—CR^(7j)R^(7k)— or—CR^(7l)R^(7m)—CR^(7n)R^(7o)C(O)—N(R^(7p))—; where each of R^(7a),R^(7b), R^(7c), R^(7d), R^(7e), R^(7f)R^(7g), R^(7h), R^(7i), R^(7j),R^(7k), R^(7l), R^(7m), R^(7n), R^(7o) and R^(7p) is independentlyselected from hydrogen and (1-4C)alkyl; each R^(8a) and R^(8b) isindependently selected from hydrogen, (1-4C)alkyl, hydroxy and fluoro,or R^(8a) and R^(8b) are joined together with the atoms to which theyare attached to form a (3-6C)cycloalkylene ring or a (2-5C)heterocyclenering containing 1 or 2 heteroatoms independently selected from oxygen,nitrogen and sulfur, or R^(8a) and W^(a) are joined together with theatoms to which they are attached to form a (2-6C)azacycloalkylene group;a is 0 or an integer of from 1 to 3; b is an integer of from 2 to 8; ora pharmaceutically acceptable salt or stereoisomer thereof.
 2. Thecompound of claim 1, wherein R¹ is an unsubstituted or substituted arylor heteroaryl group selected from a phenyl, naphthalene, pyrrole,imidazole, thiazole, oxazole, furan, thiophene, triazole, pyrazole,isoxazole, isothiazole, pyridine, pyridine N-oxide, pyrazine,pyridazine, pyrimidine, triazine, indole, benzofuran, benzothiophene,benzimidazole, benzthiazole, quinoline, isoquinoline, quinazoline andquinoxaline ring, where the point of attachment is at any availablecarbon or nitrogen ring atom.
 3. The compound of claim 2, wherein R¹ isan unsubstituted or substituted phenyl group.
 4. The compound of claim2, wherein R¹ is an unsubstituted or substituted thienyl group.
 5. Thecompound of claim 2, wherein R¹ is an unsubstituted or substitutedthiazole group.
 6. The compound of claim 2, wherein R¹ is anunsubstituted or substituted pyridyl or pyridyl N-oxide group.
 7. Thecompound of claim 1, wherein W is O.
 8. The compound of claim 1, whereinb is 2, 3 or
 4. 9. The compound of claim 1, wherein b is 3, and R^(8a)and R^(8b) on the —CR^(8a)R^(8b)— unit attached to W are methyl, and theremaining R^(8a) and R^(8b) groups are hydrogen.
 10. The compound ofclaim 1, wherein R³ is methyl.
 11. The compound of claim 1, wherein R⁶and R⁷ together form —NHC(O)CH═CH—, —CH═CHC(O)NH—, —CH₂CH₂C(O)NH— or—NHC(O)CH₂CH₂—.
 12. The compound of claim 11, wherein R⁶ and R⁷ togetherform —NHC(O)CH═CH—.
 13. The compound of claim 1, wherein the number ofcontiguous atoms in the shortest chain between the two nitrogen atoms towhich R⁴ is attached is in the range of from 8 to
 14. 14. The compoundof claim 13, wherein the number of contiguous atoms in the shortestchain between the two nitrogen atoms to which R⁴ is attached is 8, 9, 10or
 11. 15. The compound of claim 1, wherein R⁴ is a divalent group ofthe formula:—(R^(4a))_(d)-(A¹)_(e)-(R^(4b))_(f)-Q-(R^(4c))_(g)-(A²)_(h)-(R^(4d))_(i)—wherein d, e, f, g, h and i are each independently selected from 0 and1; R^(4a), R^(4b), R^(4c) and R^(d) are each independently selected from(1-10C)alkylene, (2-10C)alkenylene and (2-10C)alkynylene, wherein eachalkylene, alkenylene or alkynylene group is unsubstituted or substitutedwith from 1 to 5 substituents independently selected from (1-4C)alkyl,fluoro, hydroxy, phenyl and phenyl-(1-4C)alkyl; or R^(4d) represents(1-6C)alkylene-NHC(O)-(1-6C)alkylene; A¹ and A² are each independentlyselected from (3-7C)cycloalkylene, (6-10C)arylene, —O-(6-10C)arylene,(6-10C)arylene-O—, (2-9C)heteroarylene, —O-(2-9C)heteroarylene,(2-9C)heteroarylene-O— and (3-6C)heterocyclene, wherein eachcycloalkylene is unsubstituted or substituted with from 1 to 4substituents selected independently from (1-4C)alkyl, and each arylene,heteroarylene or heterocyclene group is unsubstituted or substitutedwith from 1 to 4 substituents independently selected from halo,(1-4C)alkyl, (1-4C)alkoxy, —S-(1-4C)alkyl, —S(O)-(1-4C)alkyl,—S(O)₂-(1-4C)alkyl, —C(O)O(1-4C)alkyl, carboxy, cyano, hydroxy, nitro,trifluoromethyl and trifluoromethoxy; Q is selected from a bond, —O—,—C(O)O—, —OC(O)—, —S—, —S(O)—, —S(O)₂—, —N(Q^(a))C(O)—, —C(O)N(Q^(b))—,—N(Q^(c))S(O)₂—, —S(O)₂N(Q^(d))—, —N(Q^(e))C(O)N(Q^(f))—,—N(Q^(g))S(O)₂N(Q^(h))—, —OC(O)N(Q^(i))—, —N(Q^(j))C(O)O— and —N(Q^(k));where Q^(a), Q^(b), Q^(c), Q^(d), Q^(e), Q^(f), Q^(g), Q^(h), Q^(i),Q^(j) and Q^(k) are each independently selected from hydrogen,(1-6C)alkyl, A³ and (1-4C)alkylene-A⁴, wherein the alkyl group isunsubstituted or substituted with from 1 to 3 substituents independentlyselected from fluoro, hydroxy and (1-4C)alkoxy; or together with thenitrogen atom and the group R^(4b) or R^(4c) to which they are attached,form a 4 to 6 membered azacycloalkylene group; and A³ and A⁴ are eachindependently selected from (3-6C)cycloalkyl, (6-10C)aryl,(2-9C)heteroaryl and (3-6C)heterocyclyl, wherein each cycloalkyl isunsubstituted or substituted with from 1 to 4 substituents selectedindependently from (1-4C)alkyl and each aryl, heteroaryl or heterocyclylgroup is unsubstituted or substituted with from 1 to 4 substituentsindependently selected from halo, (1-4C)alkyl and (1-4C)alkoxy.
 16. Thecompound of claim 15, wherein R⁴ is a divalent group of the formula:—(R^(4a))_(d)— where R^(4a) is (4-10C)alkylene.
 17. The compound ofclaim 16, wherein R⁴ is —(CH₂)₈—, —(CH₂)₉, or —(CH₂)₁₀—.
 18. Thecompound of claim 15, wherein R⁴ is a divalent group of the formula:—(R^(4a))_(d)-(A²)_(h)-(R^(4d))_(i)— wherein R^(4a) is (1-10C)alkylene;A² is (6-10C)arylene or (2-9C)heteroarylene; and R^(4d) is(1-10C)alkylene.
 19. The compound of claim 15, wherein R⁴ is a divalentgroup of the formula:—(R^(4a))_(d)-Q-(A²)_(h)-(R^(4d))_(i)— wherein Q is —O— or —N(Q^(k))-;Q^(k) is hydrogen or (1-3C)alkyl; R^(4a) is (1-10C)alkylene; A² is(6-10C)arylene or (2-9C)heteroarylene; and R^(4d) is (1-10C)alkylene.20. The compound of claim 15, wherein Q is —N(Q^(a))C(O)— or—C(O)N(Q^(b))-.
 21. The compound of claim 20, wherein R⁴ is selectedfrom:

where za is an integer from 2 to 10; and zb is an integer from 2 to 10;provided that za+zb is an integer from 4 to 12;

where zc is an integer from 2 to 7; and zd is an integer from 1 to 6;provided that zc+zd is an integer from 3 to 8; and wherein thephen-1,4-ylene group is unsubstituted or substituted with from 1 to 4substituents independently selected from halo, (1-4C)alkyl,(1-4C)alkoxy, —S-(1-4C)alkyl, —S(O)-(1-4C)alkyl, —S(O)2-(1-4C)alkyl,—C(O)O(1-4C)alkyl, carboxy, cyano, hydroxy, nitro, trifluoromethyl andtrifluoromethoxy;

where ze is an integer from 2 to 6; zf is an integer from 1 to 5; and zgis an integer from 1 to 5; provided that ze+zf+zg is an integer from 4to 8; and wherein the phen-1,4-ylene group is unsubstituted orsubstituted with from 1 to 4 substituents independently selected fromhalo, (1-4C)alkyl, (1-4C)alkoxy, —S-(1-4C)alkyl, —S(O)-(1-4C)alkyl,—S(O)₂-(1-4C)alkyl, —C(O)O(1-4C)alkyl, carboxy, cyano, hydroxy, nitro,trifluoromethyl and trifluoromethoxy;

where zh is an integer from 2 to 10; and zi is an integer from 2 to 10;provided that zh+zi is an integer from 4 to 12;

where zj is an integer from 2 to 7; and zk is an integer from 1 to 6;provided that zj+zk is an integer from 3 to 8; and wherein thephen-1,4-ylene group is unsubstituted or substituted with from 1 to 4substituents independently selected from halo, (1-4C)alkyl,(1-4C)alkoxy, —S-(1-4C)alkyl, —S(O)-(1-4C)alkyl, —S(O)₂-(1-4C)alkyl,—C(O)O(1-4C)alkyl, carboxy, cyano, hydroxy, nitro, trifluoromethyl andtrifluoromethoxy; and

where zl is an integer from 2 to 6; zm is an integer from 1 to 5; and znis an integer from 1 to 5; provided that zl+zm+zn is an integer from 4to 8; and wherein the phen-1,4-ylene group is unsubstituted orsubstituted with from 1 to 4 substituents independently selected fromhalo, (1-4C)alkyl, (1-4C)alkoxy, —S-(1-4C)alkyl, —S(O)-(1-4C)alkyl,—S(O)₂-(1-4C)alkyl, -C(O)O(1-4C)alkyl, carboxy, cyano, hydroxy, nitro,trifluoromethyl and trifluoromethoxy.
 22. The compound of claim 1,wherein R⁴ is selected from: —(CH₂)₇—; —(CH₂)₈—; —(CH₂)₉—; —(CH₂)₁₀—;—(CH₂)₁₁—; —(CH₂)₂C(O)NH(CH₂)₅—; —(CH₂)₂N(CH₃)C(O)(CH₂)₅—;—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂—; —(CH₂)₂NHC(O)(phen-1,4-ylene)CH₂—;—(CH₂)₂NHC(O)NH(CH₂)₅—; —(CH₂)₃NHC(O)NH(CH₂)₅—;—(CH₂)₂C(O)NHCH₂(cyclohex-1,3-ylene)CH₂—;—(CH₂)₂NHC(O)(cyclopent-1,3-ylene)-;—(CH₂)₂NHC(O)NH(phen-1,4-ylene)(CH₂)₂—;1-[-(CH₂)₂C(O)](piperidin-4-yl)(CH₂)₂—;—(CH₂)₂NHC(O)(trans-cyclohex-1,4-ylene)CH₂—;—(CH₂)₂NHC(O)(cis-cyclopent-1,3-ylene)-;—(CH₂)₂NH(phen-1,4-ylene)(CH₂)₂—;1-[-(CH₂)₂NHC(O)](piperidin-4-yl)(CH₂)₂—;—CH₂(phen-1,4-ylene)NH(phen-1,4-ylene)CH₂—;—(CH₂)₂C(O)NHCH₂(phen-1,3-ylene)CH₂—;—(CH₂)₂C(O)NHCH₂(pyrid-2,6-ylene)CH₂—;—(CH₂)₂C(O)NH(cis-cyclohex-1,4-ylene)CH₂—;—(CH₂)₂C(O)NH(trans-cyclohex-1,4-ylene)CH₂—;—(CH₂)₂NHC(O)(cis-cyclopent-1,3-ylene)CH₂—;—(CH₂)₂N(CH₃)C(O)(phen-1,3-ylene)CH₂—;—(CH₂)₂N(CH₃)C(O)(trans-cyclohex-1,4-ylene)CH₂—;—(CH₂)₂C(O)NH(phen-1,4-ylene)C*H(CH₃)—((S)-isomer);—(CH₂)₂C(O)NH(phen-1,4-ylene)C*H(CH₃)—((R)-isomer);2-[(S)—(—CH₂-](pyrrolidin-1-yl)C(O)(CH₂)₄—;2-[(S)-(—CH₂-](pyrrolidin-1-yl)C(O)(phen-1,4-ylene)CH₂—;—(CH₂)₂C(O)NH(4-chlorophen-1,3-ylene)CH₂—;—CH₂(2-fluorophen-1,3-ylene)CH₂—;—(CH₂)₂C(O)NH(4-methylphen-1,3-ylene)CH₂—;—(CH₂)₂C(O)NH(6-chlorophen-1,3-ylene)CH₂—;—(CH₂)₂C(O)NH(2-chlorophen-1,4-ylene)CH₂—;—(CH₂)₂C(O)NH(2,6-dichlorophen-1,4-ylene)CH₂—;—(CH₂)₂NHC(O)NHCH₂(phen-1,3-ylene)CH₂—;4-[-CH₂-](piperidin-1-yl)C(O)(phen-1,4-ylene)CH₂—;—(CH₂)₂C(O)N(CH₂CH₃)(phen-1,4-ylene)CH₂—;1-[-(CH₂)₂NHC(O)](piperidin-4-yl)-;—(CH₂)₂C(O)NH(phen-1,4-ylene)(CH₂)₂—;—(CH₂)₂NHC(O)(thien-2,5-ylene)CH₂—;—(CH₂)₂N(CH₃)C(O)(3-nitrophen-1,4-ylene)CH₂—;—(CH₂)₂N(CH₃)C(O)(trans-cyclohex-1,4-ylene)-;1-[-CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)-;5-[-(CH₂)₂NHC(O)](pyrid-2-yl)CH₂—; —(CH₂)₂(phen-1,4-ylene)(CH₂)₂—;—(CH₂)₃(thien-2,5-ylene)(CH₂)₃—;—(CH₂)₂(phen-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂—;—CH₂(phen-1,2-ylene)NH(phen-1,4-ylene)(CH₂)₂—;1-[-CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)(CH₂)₂—;1-[-CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)CH₂—;—(CH₂)₂C(O)NH(3-chlorophen-1,4-ylene)CH₂—;—(CH₂)₂C(O)NH(2-(CF₃O-)phen-1,4-ylene)CH₂—;—(CH₂)₃(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂—; —(CH₂)₂S(O)₂NH(CH₂)₅—;—CH₂(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂—;—(CH₂)₂C(O)NH(2-iodophen-1,4-ylene)CH₂—;—(CH₂)₂C(O)NH(2-chloro-5-methoxyphen-1,4-ylene)CH₂—;—(CH₂)₂C(O)NH(2-chloro-6-methylphen-1,4-ylene)CH₂—;—(CH₂)₂N(CH₃)S(O)₂(phen-1,4-ylene)CH₂—;—(CH₂)₂C(O)NH(2-bromophen-1,4-ylene)CH₂—;—(CH₂)₃(phen-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂—;—(CH₂)₃(phen-1,2-ylene)NH(phen-1,4-ylene)(CH₂)₂—;1-[-CH₂(2-fluorophen-1,3-ylene)CH₂](piperidin-4-yl)(CH₂)₃—;—(CH₂)₂C(O)NH(2-methoxyphen-1,4-ylene)CH₂—;—(CH₂)₅NH(phen-1,4-ylene)(CH₂)₂—;4-[-(CH₂)₂-](piperidin-1-yl)(phen-1,4-ylene)(CH₂)₂—;—(CH₂)₂C(O)NH(phen-1,4-ylene)CH(CH₃)CH₂—;—(CH₂)₂-(trans-cyclohex-1,4-ylene)NH(phen-1,4-ylene)(CH₂)₂—;—(CH₂)₂C(O)NH(2-fluorophen-1,4-ylene)CH₂—;—(CH₂)₂(phen-1,3-ylene)NH(phen-1,4-ylene)(CH₂)₂—;—(CH₂)₂C(O)NH(2,5-difluorophen-1,4-ylene)CH₂—;—(CH₂)₂NHC(O)(phen-1,4-ylene)(CH₂)₂—;1-[—CH₂(pyrid-2,6-ylene)CH₂](piperidin-4-yl)CH₂—;—(CH₂)₃NH(phen-1,4-ylene)(CH₂)₂—; —(CH₂)₂NH(naphth-1,4-ylene)(CH₂)₂—;—(CH₂)₃O(phen-1,4-ylene)CH₂—; 1-[-(CH₂)₃](piperidin-4-yl)CH₂—;4-[-(CH₂)₂](piperidin-1-yl)C(O)(phen-1,4-ylene)CH₂—;—(CH₂)₃(phen-1,4-ylene)NHC(O)(CH₂)₂—; —(CH₂)₃O(phen-1,4-ylene)(CH₂)₂—;2-[-(CH₂)₂](benzimidazol-5-yl)CH₂—;—(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₂—;—(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₄—;—(CH₂)₂-(trans-cyclohex-1,4-ylene)NHC(O)(CH₂)₅—;4-[-(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₂—;—(CH₂)₂NHC(O)NH(phen-1,4-ylene)CH₂—;—(CH₂)₂N(CH₃)(CH₂)₂(cis-cyclohex-1,4-ylene)-;—(CH₂)₂C(O)NH(2,3,5,6-tetrafluorophen-1,4-ylene)CH₂—;—(CH₂)₂C(O)NH(2,6-diiodophen-1,4-ylene)CH₂—;4-[-(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₃—;4-[-(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₄—;4-[-(CH₂)₂](piperidin-1-yl)C(O)(CH₂)₅—;—(CH₂)₂C(O)NHCH₂(phen-1,4-ylene)CH₂—;—(CH₂)₂NHC(O)NHCH₂(phen-1,4-ylene)CH₂—;—(CH₂)₂C(O)NH(2-methylphen-1,4-ylene)CH₂—;1-[-(CH₂)₃O(phen-1,4-ylene)(CH₂)₂](piperidin-4-yl)CH₂—;—(CH₂)₂C(O)NHCH₂(phen-1,3-ylene)(CH₂)₂—; —(CH₂)₂O(phen-1,3-ylene)CH₂—;—(CH₂)₂N(CH₃)C(O)CH₂O(phen-1,4-ylene)CH₂—;—(CH₂)₂N(CH₃)C(O)CH₂O(phen-1,3-ylene)CH₂—;—(CH₂)₂N(CH₃)C(O)(fur-2,5-ylene)CH₂—;—(CH₂)₂N(CH₃)C(O)(thien-2,5-ylene)CH₂—;—(CH₂)₂O(phen-1,4-ylene)O(CH₂)₂—;—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(phen-1,4-ylene)CH₂—;—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)CH₂O(phen-1,2-ylene)CH₂—;—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)CH₂O(phen-1,3-ylene)CH₂—;—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)CH₂O(phen-1,4-ylene)CH₂—;—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(fur-2,5-ylene)CH₂—;—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(thien-2,5-ylene)CH₂—;4-[-(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,2-ylene)CH₂—;4-[-(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,3-ylene)CH₂—;4-[-(CH₂)₂](piperidin-1-yl)C(O)CH₂O(phen-1,4-ylene)CH₂—;4-[-(CH₂)₂](piperidin-1-yl)C(O)(fur-2,5-ylene)CH₂—;4-[-(CH₂)₂](piperidin-1-yl)C(O)(thien-2,5-ylene)CH₂—;—(CH₂)₂(phen-1,4-ylene)NHC(O)(phen-1,3-ylene)CH₂—;—(CH₂)₂(phen-1,4-ylene)NHC(O)(phen-1,4-ylene)CH₂—;—(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,2-ylene)CH₂—;—(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,3-ylene)CH₂—;—(CH₂)₂(phen-1,4-ylene)NHC(O)CH₂O(phen-1,4-ylene)CH₂—;—(CH₂)₂(phen-1,4-ylene)NHC(O)(fur-2,5-ylene)CH₂—;—(CH₂)₂(phen-1,4-ylene)NHC(O)(thien-2,5-ylene)CH₂—;—(CH₂)₂(trans-cyclohex-1,4-ylene)NHC(O)(phen-1,3-ylene)CH₂—;—(CH₂)₃O(phen-1,3-ylene)CH₂—; —CH₂CH(OH)CH₂NH(phen-1,4-ylene)(CH₂)₂—;—(CH₂)₄NH(phen-1,4-ylene)(CH₂)₂—;—(CH₂)₂C(O)NH(phen-1,4-ylene)CH₂NHC(O)CH₂—;—(CH₂)₂C(O)NH(phen-1,4-ylene)(CH₂)₂NHC(O)CH₂—;—(CH₂)₂C(O)NHCH₂(trans-cyclohex-1,4-ylene)CH₂—; —(CH₂)₂NHC(O)(CH₂)₅—;—(CH₂)₂O(phen-1,3-ylene)O(CH₂)₂—; —(CH₂)₂O(phen-1,2-ylene)O(CH₂)₂—;—CH₂(phen-1,2-ylene)O(phen-1,2-ylene)CH₂—; —(CH₂)₂C(O)NH(CH₂)₆—;—(CH₂)₃(phen-1,4-ylene)(CH₂)₃—; —(CH₂)₃(phen-1,4-ylene)(CH₂)₂—;—(CH₂)₄(phen-1,4-ylene)(CH₂)₂—; —(CH₂)₃(furan-2,5-ylene)(CH₂)₃—;—(CH₂)₂N(CH₃)C(O)NH(phen-1,4-ylene)(CH₂)₂—;4-[-(CH₂)₂](piperidin-1-yl)C(O)NH(phen-1,4-ylene)(CH₂)₂—;—(CH₂)₃(phen-1,3-ylene)(CH₂)₃—;—(CH₂)₃(tetrahydrofuran-2,5-ylene)(CH₂)₃—; and—(CH₂)₂O(phen-1,4-ylene)C(O)(CH₂)₂—.
 23. The compound of claim 1,wherein the compound is a compound of formula II:

or a pharmaceutically acceptable salt or stereoisomer thereof.
 24. Thecompound of claim 1, wherein the compound is a compound of formula V:

wherein each R⁹ is independently selected from (1-4C)alkyl,(2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(9a),—C(O)OR^(9b), —SR^(9c), —S(O)R^(9d), —S(O)₂R^(9e) and —NR^(9f)R^(9g);wherein R^(9a), R^(9b), R^(9c), R^(9d), R^(9e), R^(9f) and R^(9g) areindependently selected from hydrogen, (1-4C)alkyl andphenyl-(1-4C)alkyl; and n is 0, 1, 2 or 3; or a pharmaceuticallyacceptable salt or stereoisomer thereof.
 25. The compound of claim 1,wherein the compound is a compound of formula VI:

wherein each R¹⁰ is independently selected from (1-4C)alkyl,(2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(10a),—C(O)OR^(10b), —SR^(10c), —S(O)R^(10d), —S(O)₂R^(10e) and—NR^(10f)R^(10g); wherein R^(10a), R^(10b), R^(10c), R^(10d), R^(10e),R^(10f) and R^(10g) are independently selected from hydrogen,(1-4C)alkyl and phenyl-(1-4C)alkyl; and o is 0, 1, 2 or 3; or apharmaceutically acceptable salt or stereoisomer thereof.
 26. Thecompound of claim 1, wherein the compound is a compound of formula VII:

wherein each R¹¹ is independently selected from (1-4C)alkyl,(2-4C)alkenyl, (2-4C)alkynyl, (3-6C)cycloalkyl, cyano, halo, —OR^(11a),—C(O)OR^(11b), —SR^(11c), —S(O)R^(11d), —S(O)₂R^(11e) and—NR^(11f)R^(11g); wherein R^(11a), R^(11b), R^(11c), R^(11d), ^(11e),R^(11f) and R^(11g) are independently selected from hydrogen,(1-4C)alkyl or phenyl-(1-4C)alkyl; and p is 0, 1 or 2; or apharmaceutically acceptable salt or stereoisomer thereof.
 27. Thecompound of claim 1, wherein the compound is a compound of formula VIII:

or the corresponding pyridine N-oxide, wherein each R¹² is independentlyselected from (1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl,(3-6C)cycloalkyl, cyano, halo, —OR^(12a), —C(O)OR^(12b), —SR^(12c),—S(O)R^(12d), —S(O)₂R^(12e) and —NR^(12f)R^(12g); wherein R^(12a),R^(12b), R^(12c), R^(12d), R^(12e), R^(12f) and R^(12g) areindependently selected from hydrogen, (1-4C)alkyl andphenyl-(1-4C)alkyl; and q is 0, 1 or 2; or a pharmaceutically acceptablesalt or stereoisomer thereof.
 28. The compound of claim 1, wherein thecompound is selected from: biphenyl-2-ylcarbamic acid3-({9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydro-quinolin-5-yl)ethylamino]nonyl}methylamino)-1,1-dimethylpropylester; (2-thien-3-ylphenyl)carbamic acid3-({9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]nonyl}methylamino)-1,1-dimethylpropylester; biphenyl-2-ylcarbamic acid3-{[2-(4-{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}benzoylamino)ethyl]methylamino}-1,1-dimethylpropylester; biphenyl-2-ylcarbamic acid3-{[4-(4-{2-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]ethyl}phenyl)butyl]methylamino}-1,1-dimethyl-propylester; and biphenyl-2-ylcarbamic acid3-{[2-(2-chloro-4-{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydroquinolin-5-yl)ethylamino]methyl}-5-methoxyphenylcarbamoyl)ethyl]-methylamino}-1,1-dimethylpropylester.
 29. A pharmaceutical composition comprising a pharmaceuticallyacceptable carrier and a therapeutically effective amount of a compoundof claim
 1. 30. The pharmaceutical composition of claim 29, wherein thecomposition further comprises a therapeutically effective amount of asteroidal anti-inflammatory agent.
 31. The pharmaceutical composition ofclaim 29, wherein the composition further comprises a therapeuticallyeffective amount of a PDE₄ inhibitor.
 32. A process for preparing acompound of claim 1, the process comprising: (a) reacting a compound offormula 1:

or a salt thereof; with a compound of formula 2:

wherein X¹ represents a leaving group, and P¹ and P² each independentlyrepresent a hydrogen atom or a hydroxyl-protecting group; (b) reacting acompound of formula 3:

or salt thereof; with a compound of formula 4:

wherein X² represents a leaving group, and P³ and P⁴ each independentlyrepresent a hydrogen atom or a hydroxyl-protecting group; (c) coupling acompound of formula 5:

with a compound of formula 6:

wherein X^(Qa) and X^(Qb) each independently represent functional groupsthat couple to form a group Q, P^(5a) represents a hydrogen atom or anamino-protecting group; and P^(5b) and P⁶ each independently represent ahydrogen atom or a hydroxyl-protecting group; (d) for a compound offormula I wherein R⁵ represents a hydrogen atom, reacting a compound offormula 3 with a compound of formula 7:

or a hydrate thereof (e.g., a glyoxal), in the presence of a reducingagent, wherein P⁷ represents a hydrogen atom or a hydroxyl-protectinggroup; (e) reacting a compound of formula I with a compound of formula8:

or a hydrate thereof, in the presence of a reducing agent, wherein P⁸and P⁹ each independently represent a hydrogen atom or ahydroxyl-protecting group, P¹⁰ represents a hydrogen atom or anamino-protecting group, and R⁴ represents a residue that, together withthe carbon to which it is attached, affords a group R⁴ upon completionof the reaction; (f) reacting a compound of formula 9:

wherein X³ represents a leaving group, with a compound of formula 10:

wherein P¹¹ and P¹² each independently represent a hydrogen atom or ahydroxyl-protecting group, and P¹³ represents a hydrogen atom or anamino-protecting group; or (g) reacting a compound of formula 11:

or a hydrate thereof; wherein R⁴ represents a residue that, togetherwith the carbon to which it is attached, affords a group R⁴ uponcompletion of the reaction; with a compound of formula 10 in thepresence of a reducing agent; (h) reacting a compound of formula 12:

wherein Y¹ represents chloro, bromo, iodo or CF₃SO₂O—, P¹⁴ represents ahydrogen atom or an amino-protecting group; and P¹⁵ and P¹⁶ eachindependently represent a hydrogen atom or a hydroxyl-protecting group;with a compound of the formula: R¹—B(OH)₂ in the presence of a couplingcatalyst; and then removing any protecting group P¹, P², P³, P⁴, P^(5a),P^(5b), P⁶, P⁷, P⁸, P⁹, P¹⁰, P¹¹, P¹², P¹³, P¹⁴, P¹⁵ and P¹⁶ to providea compound of formula I, or a salt thereof.
 33. The process of claim 32,wherein the process further comprises forming a pharmaceuticallyacceptable salt of the compound of formula I.
 34. A compound of formula12:

wherein Y¹ represents chloro, bromo, iodo or CF₃SO₂O—; W represents O orNW^(a); where W^(a) is hydrogen or (1-4C)alkyl; each R² is independentlyselected from (1-4C)alkyl, (2-4C)alkenyl, (2-4C)alkynyl,(3-6C)cycloalkyl, cyano, halo, —OR^(2a), —C(O)OR^(2b), —SR^(2c),—S(O)R^(2d), —S(O)₂R^(2c) and —NR^(2f)R^(2g); where each of R^(2a),R^(2b), R^(2c), R^(2d), R^(2e), R^(2f) and R^(2g) is independentlyselected from hydrogen, (1-4C)alkyl or phenyl-(1-4C)alkyl; and whereeach alkyl group present in R² is unsubstituted or substituted with from1 to 3 fluoro substituents; R³ is selected from hydrogen, (1-6C)alkyl,(2-6C)alkenyl, (2-6C)alkynyl, or (3-6C)cycloalkyl, where the alkyl groupis unsubstituted or substituted with from 1 to 3 fluoro substituents; R⁴represents a divalent hydrocarbon group containing from 4 to 28 carbonatoms and optionally containing from 1 to 10 heteroatoms selectedindependently from halo, oxygen, nitrogen and sulfur, provided that thenumber of contiguous atoms in the shortest chain between the twonitrogen atoms to which R⁴ is attached is in the range of from 4 to 16;R⁵ represents hydrogen or (1-4C)alkyl; R⁶ and R⁷ together form—N(R^(7a))C(O)C(R^(7b))═C(R^(7c))—, —C(R^(7d))═C(R^(7c))C(O)N(R^(7f))—,—N(R^(7g))C(O)CR^(7h)R^(7i)—CR^(7j)R^(7k)— or—CR^(7l)R^(7m)—CR^(7n)R^(7o)C(O)—N(R^(7p))—; where each of R^(7a),R^(7b), R^(7c), R^(7d), R^(7e), R^(7f)R^(7g), R^(7h), R^(7i), R^(7j),R^(7k), R^(7l), R^(7m), R^(7n), R^(7o) and R^(7p) is independentlyselected from hydrogen and (1-4C)alkyl; each R^(8a) and R^(8b) isindependently selected from hydrogen and (1-4C)alkyl, or R^(8a) andR^(8b) are joined together with the atoms to which they are attached toform a (3-6C)cycloalkylene ring or a (2-5C)heterocyclene ring containing1 or 2 heteroatoms independently selected from oxygen, nitrogen andsulfur, or R^(8a) and W^(a) are joined together with the atoms to whichthey are attached to form a (2-6C)azacycloalkylene group; P¹⁴ representsa hydrogen atom or an amino-protecting group; P¹⁵ and P¹⁶ eachindependently represent a hydrogen atom or a hydroxyl-protecting group;a is 0 or an integer of from 1 to 3; b is an integer of from 2 to 8; ora salt or stereoisomer thereof.
 35. The compound of claim 34, wherein Y¹is bromo.